CN116380867A - Raman/fluorescence dual-light-path spectrum detection device and detection method - Google Patents

Abstract

The invention discloses a Raman/fluorescence dual optical path spectrum detection device and detection method. By setting a sample on a confocal microscope, when an excitation light source controlled by an optical switch is connected to the optical path, according to its use, Connect the corresponding filter structure to the optical path, the excitation light output by the laser light source will sequentially pass through the first mirror, filter structure and confocal microscope, and then irradiate the sample, and generate signal light, which is Raman light Or fluorescence, and then pass the signal light through the filter structure to filter out unnecessary laser signals and then enter the monochromator through the reflector and focusing mirror to obtain the spectrum. Among them, by setting an optical switch, it can be used to emit two kinds of excitation light sources with different wavelengths; by setting a filter structure, different light can be obtained; by setting a confocal microscope, it can be used to reflect and collect light. Therefore, the device realizes the effective integration of the Raman spectrometer and the fluorescence spectrometer, and the acquisition of the Raman/fluorescence spectrum.

Description

A kind of Raman/fluorescence dual optical path spectrum detection device and detection method

technical field

The invention belongs to the technical field of instrument testing, and relates to a Raman/fluorescence dual optical path spectrum detection device and a detection method.

Background technique

Fluorescence refers to a luminescence phenomenon called photoluminescence. A substance at normal temperature is irradiated by incident light of a certain wavelength, that is, the usual ultraviolet light, absorbs light energy and enters an excited state, then immediately returns to the ground state, and emits outgoing light with a longer wavelength than the incident light. Usually the wavelength is in the visible light band. Raman spectroscopy is based on the inelastic scattering of radiation by a sample, which can be a solid, liquid or gas. In practice, when incident monochromatic light interacts with molecules, there are two types of light scattering (elastic and inelastic). During elastic scattering (Rayleigh scattering), there is no change in the frequency of the photon, while inelastic scattering (Raman scattering), there is an accompanying change in the frequency of the photon. The frequency difference between the scattered photon and the incident photon is the Raman shift (cm ^-1 ), which includes the vibration information related to the molecule.

As the most widely used detection methods, the advantages and disadvantages of fluorescence and Raman spectroscopy are highly complementary. Fluorescence spectroscopy is the most sensitive and fastest method for detecting organic substances and biomarkers. Raman spectroscopy has high resolution and specificity, and there will be fluorescence signals during Raman detection, which are only filtered out as background noise. Therefore, it is desirable to collect the fluorescence signal generated during Raman spectroscopy measurement and perform fluorescence detection and Raman detection simultaneously. In addition, the combination of fluorescence spectroscopy and Raman spectroscopy can combine the advantages of the two to obtain a detection method with strong specificity and high sensitivity.

However, due to the essential differences in the realization principles of Raman spectroscopy and fluorescence spectroscopy, the internal structures of Raman spectrometers and fluorescence spectrometers are significantly different. There are technical difficulties in combining the two into a Raman-fluorescence combined spectrometer: two The wavelengths of the lasers used by the latter are different; the types of dichroic mirrors used to separate the excitation light from the generated signal light required by the two are different; in addition, due to the different spectral ranges of Raman and fluorescence, when the spectrum is split The type of grating required is different. The existence of these problems makes it difficult to integrate a Raman spectrometer and a fluorescence spectrometer into one spectrometer.

Contents of the invention

The purpose of the present invention is to solve the problem in the prior art that it is difficult to combine the two to obtain Raman spectrum or fluorescence spectrum due to the significant difference in the internal structure of Raman spectrometer and fluorescence spectrometer, and to provide a Raman/fluorescence dual optical path Spectrum detection device and detection method.

In order to achieve the above object, the present invention adopts the following technical solutions to achieve:

A Raman/fluorescence dual optical path spectrum detection device proposed by the present invention includes an optical switch, a first mirror, a confocal microscope, a focusing mirror, a third mirror, a filter structure and a monochromator;

A sample is set on the confocal microscope; the first laser or the second laser light is irradiated on the sample after sequentially passing through the optical switch, the first mirror, the filter structure and the confocal microscope, and the light excited in the sample is sequentially After passing through the confocal microscope, filter structure, third mirror and focusing mirror into the monochromator, the spectrum is obtained.

Preferably, a neutral filter is provided between the optical switch and the first reflector; a confocal beam expander is provided between the first reflector and the filter structure.

Preferably, the filter structure includes a Raman filter set, a fluorescence filter set and a filter wheel, and the Raman filter set and the fluorescence filter set are arranged on the filter on the sheet wheel; the Raman filter set includes a first long-pass dichroic mirror and a first notch filter, and the fluorescence filter set includes a second long-pass dichroic mirror and a second Notch filter;

After the first laser light passes through the confocal beam expander, it is switched to a Raman filter group through the filter wheel, and the first long-pass dichroic mirror reflects the first laser light to the confocal microscope, and then receives the confocal The wavelength of the Raman light from the microscope is greater than that of the first laser, and the first laser is filtered out by the first notch filter;

Or, after the second laser light passes through the confocal beam expander, it is switched to a fluorescence filter group through the filter wheel, and the second long-pass dichroic mirror reflects the second laser light to the confocal microscope, and then receives the confocal The wavelength transmitted by the focusing microscope is greater than the fluorescence of the second laser light, and then the second laser light is filtered out by the second notch filter.

Preferably, the exteriors of the Raman filter set, the fluorescence filter set, the filter wheel, the reflection mirror and the focusing mirror are packaged in a black interface box.

Preferably, the confocal microscope includes a second mirror, a dual-port optical path switching device, a microscope objective lens, and a two-dimensional displacement stage; the dual-port optical path switching device includes a first displacement mechanism, a first position limiter and a semi-transparent A half mirror; the half mirror and the first position limiter are both arranged on the first displacement structure; the two-dimensional displacement stage is vertically arranged on the plane of the main optical axis of the confocal microscope;

After the Raman light passes through the second mirror, the half-mirror is moved out of the optical path. After the two-dimensional displacement stage drives the sample to receive the Raman light irradiation, the excited Raman light passes through the first long-pass dichroic The mirror, the first notch filter, the mirror and the focusing mirror are sent to the monochromator to obtain the Raman spectrum;

Or, after the fluorescence passes through the second reflector, move the half-mirror out of the optical path, and after the sample is driven by the two-dimensional displacement stage to receive the fluorescence irradiation, the excited fluorescence passes through the second long-pass dichroic mirror, the second Notch filters, mirrors, and focusing mirrors are sent to a monochromator to obtain fluorescence spectra.

Preferably, a first video camera is provided on one side of the first displacement structure;

When the Raman light/fluorescence passes through the second mirror, the half-mirror is moved into the optical path, and the signal light is split by the half-mirror and enters the video camera.

Preferably, the first laser is a 633nm laser, generated by a first laser;

The second laser is 532nm laser, which is generated by the second laser, and the second laser emitted by the second laser is reflected into the neutral filter through the fourth reflector under the action of the optical switch.

Preferably, the monochromator includes a first concave reflector, a second concave reflector and a grating cone, on which a plurality of gratings for dispersing light are installed;

Raman light or fluorescence passes through the focusing mirror and passes through the first concave reflector, the grating cone wheel and the second concave reflector in sequence, to obtain the spectrum;

A photoelectric converter is connected to the output end of the second concave reflector.

Preferably, the optical switch includes a second displacement mechanism, a second position limiter and a fifth reflector; both the fifth reflector and the second position limiter are mounted on the second displacement mechanism.

A kind of Raman/fluorescence dual optical path spectral detection method proposed by the present invention comprises the following steps:

Place the sample on the confocal microscope, and connect the first laser/second laser to the optical path through an optical switch;

Connect the first mirror, confocal microscope, focusing mirror, third mirror, filter structure and monochromator into the optical path;

The first laser/second laser sequentially passes through the first reflector, filter structure and confocal microscope, and then irradiates the sample. The excited Raman light passes through the confocal microscope, filter structure, third reflector and focus in turn. After passing into the monochromator behind the mirror, the spectrum is obtained.

Compared with the prior art, the present invention has the following beneficial effects:

A Raman/fluorescence dual optical path spectrum detection device proposed by the present invention, by setting a sample on a confocal microscope, when an excitation light source controlled by an optical switch is connected to the optical path, according to its different uses, the The corresponding filter structure is connected to the optical path, and the excitation light output by the laser light source will sequentially pass through the first mirror, filter structure and confocal microscope, and then irradiate the sample to generate signal light. The signal light is Raman light or Fluorescence, and then the signal light passes through the filter structure to filter out unnecessary laser signals, and then enters the monochromator through the reflector and focusing mirror to obtain the spectrum. Among them, by setting an optical switch, it can be used to emit two kinds of excitation light sources with different wavelengths; by setting a filter structure, different light can be obtained; by setting a confocal microscope, it can be used to reflect and collect light. Based on the above statements, it is shown that the detection device proposed by the present invention realizes the effective integration of Raman spectrometer and fluorescence spectrometer, has high space utilization rate, can obtain Raman and fluorescence spectra, and has wide application in the fields of food safety, nanomaterial analysis, criminal investigation, archaeology, etc. application prospects.

Further, the operator can judge the detection site of the sample through the video camera.

Furthermore, the neutral filter can adjust the excitation light power to 25%, 50%, 75% and 100% of the initial power, avoiding the central wavelength that may be caused by adjusting the optical power through the power adjustment knob that comes with the laser. offset problem.

Further, the diameter of the beam can be adjusted by setting a confocal beam expander until the Raman/fluorescence spectrum with the best intensity is obtained.

The invention provides a Raman/fluorescence dual optical path spectrum detection method, under the action of an optical switch, the first laser or the second laser is sequentially injected into the first reflector, filter structure and confocal microscope, and then irradiated to the sample In the above, after the light excited from the sample passes through the confocal microscope, filter structure, third mirror and focusing mirror into the monochromator in sequence, the required spectrum can be obtained. This method is simple to operate and easy to implement.

Description of drawings

In order to illustrate the technical solutions of the embodiments of the present invention more clearly, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention, and thus It should be regarded as a limitation on the scope, and those skilled in the art can also obtain other related drawings based on these drawings without creative work.

Fig. 1 is a schematic diagram of the working state of the Raman/fluorescence dual optical path spectrum detection device of the present invention for Raman spectrum detection;

Fig. 2 is a schematic diagram of the working state of the Raman/fluorescence dual optical path spectrum detection device of the present invention for fluorescence spectrum detection;

Fig. 3 is a schematic diagram of the working state if the dual-port switching device controls the video camera to move out of the optical path in the example of the present invention.

Fig. 4 is a flow chart of the Raman/fluorescence dual optical path spectral detection method of the present invention.

Among them: 1-first laser; 2-second laser; 3-optical switch; 4-neutral filter; 5-first mirror; 6-confocal beam expander; 7-interface box; 8-common Focusing microscope; 9-second mirror; 10-dual-port optical path switching device; 11-video camera; 12-microscopic objective lens; 13-two-dimensional displacement platform; 14-photoelectric converter; The first concave reflector; 17-the second concave reflector; 18-focusing mirror; 19-the third reflector; 20-the first notch filter; 21-the first long-pass dichroic mirror; 22-the first Two notch filters; 23-the third long-pass dichroic mirror; 24-filter wheel, 25-the fourth reflector, 26-the fifth reflector, 27-half-transparent mirror.

Detailed ways

In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments It is a part of embodiments of the present invention, but not all embodiments. The components of the embodiments of the invention generally described and illustrated in the figures herein may be arranged and designed in a variety of different configurations.

Accordingly, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely represents selected embodiments of the invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

It should be noted that like numerals and letters denote similar items in the following figures, therefore, once an item is defined in one figure, it does not require further definition and explanation in subsequent figures.

In the description of the embodiments of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "upper", "lower", "horizontal", "inside" etc. is based on the orientation or positional relationship shown in the drawings , or the orientation or positional relationship that the product of the invention is usually placed in use is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation or be constructed in a specific orientation and operation, and therefore should not be construed as limiting the invention. In addition, the terms "first", "second", etc. are only used for distinguishing descriptions, and should not be construed as indicating or implying relative importance.

In addition, when the term "horizontal" appears, it does not mean that the part is required to be absolutely horizontal, but may be slightly inclined. For example, "horizontal" only means that its direction is more horizontal than "vertical", and it does not mean that the structure must be completely horizontal, but can be slightly inclined.

In the description of the embodiments of the present invention, it should also be noted that, unless otherwise specified and limited, the terms "setting", "installation", "connection" and "connection" should be interpreted in a broad sense, for example, It can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediary, and it can be the internal communication of two components. Those of ordinary skill in the art can understand the specific meanings of the above terms in the present invention according to specific situations.

The present invention is described in further detail below in conjunction with accompanying drawing:

The present invention provides a Raman/fluorescence dual optical path spectrum detection device, as shown in Figure 1 and Figure 2, comprising: a laser unit for emitting excitation light sources of two different wavelengths, and a light collection device for reflecting and collecting light A unit, a filter structure and a spectral detection unit for obtaining spectral information.

The laser unit includes a 633nm first laser 1 for Raman detection, a 532nm second laser 2 for fluorescence detection, an optical switch 3, a neutral filter 4, a confocal beam expander 6 and a fourth mirror 25. The optical switch 3 includes a second displacement mechanism, a second position limiter and a fifth reflection mirror 26, and the second position limiter and the fifth reflection mirror 26 are both arranged on the second displacement mechanism. Wherein, the second displacement mechanism is used to move the fifth reflecting mirror 26 into or out of the position of the main optical axis. When the fifth reflector 26 is moved out of the main optical axis, the 633nm first laser 1 is connected to the optical path; when the fifth reflector 26 is moved into the main optical axis, the 375nm second laser 2 is connected to the optical path. The beam diameter adjustment range of the confocal beam expander 6 is 1×-2×.

The light collection unit is a confocal microscope 8, on which a sample is arranged, and the confocal microscope 8 includes a second mirror 9 for reflecting and folding light, thereby improving the space utilization of the system, and a dual-port optical path switching device 10 , a microscope objective lens 12 and a two-dimensional displacement stage 13. The dual-port optical path switching device 10 includes a first displacement mechanism, a first position stopper and a half mirror 27, and the first position stopper and the half mirror 27 are all arranged on the first displacement mechanism, and the first position stopper and the half mirror 27 are all arranged on the first displacement mechanism. One side of the displacement structure is provided with a first video camera 11 . The second displacement mechanism is used to move the half-mirror 27 into or out of the working optical path. When it is moved into the working optical path, the signal light is split by the half-mirror 27 and enters the video camera 11 so that the experimenter observes. When the mirror 27 moves out of the optical path, the video camera 11 no longer receives light information, and the reflected light beams all enter the filter structure, which can avoid the problem that the light intensity is halved after passing through the half mirror 27. The two-dimensional translation stage 13 can move freely on a plane perpendicular to the main optical axis of the confocal microscope 8, and can realize fluorescence or Raman detection of multiple positions on the two-dimensional layer of the sample.

When working, after the Raman light passes through the second reflector 9, the half-transparent mirror 27 is moved out of the optical path, and the two-dimensional displacement stage 13 drives the sample to receive the Raman light irradiation, and the excited Raman light passes through the second mirror in turn. A long-pass dichroic mirror 21, the first notch filter 20, the mirror 19 and the focusing mirror 18 are sent to the monochromator to obtain the Raman spectrum;

Or, after the fluorescence passes through the second reflector 9, the half-transparent mirror 27 is moved out of the optical path, and after the two-dimensional displacement stage 13 drives the sample to receive the fluorescence irradiation, the excited fluorescence passes through the second long-pass dichroic mirror in turn 23. The second notch filter 22, the mirror 19 and the focusing mirror 18 are sent to the monochromator to obtain the fluorescence spectrum.

The filter structure includes a Raman filter group, a fluorescence filter group, a filter wheel 24, a third reflector 19 and a focusing mirror 18, and this part is packaged with a light-shielding black interface box 7; the Raman filter The set includes a first longpass dichroic mirror 21 at 633 nm and a first notch filter 20 at 633 nm. The fluorescence filter set includes a 375nm second long-pass dichroic mirror 21 and a 375nm second notch filter 20 . The first long-pass dichroic mirror 21 is used to reflect 633nm laser light, and receives Raman scattered light with a wavelength greater than 633nm, and the first notch filter 20 is used to filter out 633nm laser light; the fluorescence filter group, the second long-pass The dichroic mirror 23 is used to reflect the 375nm laser light, and receives the fluorescent signal with a wavelength greater than 375nm, and the second notch filter 22 is used to filter out the 375nm laser light; the filter wheel 24 is used to realize the Raman filter of the access optical path Manual switching of optical filter set or fluorescence filter set.

During operation, after the first laser light passes through the confocal beam expander 6, it is switched to a Raman filter group through the filter wheel 24, and the first long-pass dichroic mirror 21 reflects the first laser light to the confocal microscope 8 , and then receive the Raman light from the confocal microscope 8 with a wavelength greater than that of the first laser, and then pass through the first notch filter 20 to filter out the first laser; or, the second laser passes through the confocal beam expander 6 Finally, the filter wheel 24 is used to switch to a fluorescent filter set, and the second long-pass dichroic mirror 23 reflects the second laser light to the confocal microscope 8, and then receives the wavelength transmitted by the confocal microscope 8 that is greater than the second laser beam. The fluorescence of the laser light is filtered by the second notch filter 22 to filter out the second laser light.

The spectral detection unit includes a monochromator and a photoelectric converter 14. The monochromator includes two first concave reflectors 16, a second concave reflector 17 and a grating tower wheel 15 for folding the optical path. There are several gratings for dispersing light, and the operator can switch according to different functional requirements.

During work, Raman light or fluorescence passes through the focusing mirror 18 and passes through the first concave reflector 16, the grating tower wheel 15 and the second concave reflector 17 in sequence to obtain a spectrum, and the output end of the second concave reflector 17 is connected to a photoelectric converter 14, is to convert the spectral signal into an electrical signal for later use.

When an excitation light source controlled by an optical switch is connected to the optical path, according to the different uses, the corresponding filter group is connected to the optical path through the filter wheel 24, and the excitation light output by the laser light source will be After passing through the neutral filter 4 and the confocal beam expander 6 in turn, and then reflected by the first long-pass dichroic mirror 21/second long-pass dichroic mirror 23 in the interface box 7, it is focused by the microscope objective lens 12 to the sample and generate signal light, the signal light is Raman light or fluorescence, the signal light enters the dual-port optical path switching device 10 through the microscope objective lens 12, which contains a half-transparent mirror 27 that can split the signal light and send it to The video camera 11 is convenient for experimenters to observe, as shown in FIG. 3 . In addition, the operator can move the half-mirror 27 in the dual-port optical path switching device 10 out of the optical path according to needs, so as to avoid the problem that the light intensity is halved after passing through the half-mirror 27 . The light beam passes through the filter set in the interface box 7 , filters the laser signal, enters the monochromator through the reflector 19 and the focusing mirror 18 , and then enters the photoelectric converter 14 .

A specific Raman spectrum detection implementation is shown in FIG. 1 , and a fluorescence spectrum detection implementation is shown in FIG. 2 . The detection method of the Raman/fluorescence dual optical path spectrum detection device proposed by the present invention, as shown in Figure 4, comprises the following steps:

Step 1. Place the sample on the confocal microscope 8, and connect the first laser/second laser to the optical path through the optical switch 3;

Step 2, connecting the first mirror 5, the confocal microscope 8, the focusing mirror 18, the third mirror 19, the filter structure and the monochromator into the optical path;

Step 3. The first laser light/second laser light is irradiated onto the sample after sequentially passing through the first reflector 5, filter structure and confocal microscope 8, and the excited Raman light passes through the confocal microscope 8, filter structure, After the third reflection mirror 19 and the focusing mirror 18 are passed into the monochromator, the spectrum is obtained.

The implementation mode includes the following steps:

(1) Raman Spectroscopy Detection

a, placing a planar solid or liquid sample on the two-dimensional displacement stage 13, the sample may also contain a part for fluorescence or Raman spectrum detection;

b, connecting the Raman filter group into the optical path by rotating the filter wheel 24;

c, connecting the first 633nm laser 1 into the optical path through the optical switch 3;

d, connecting the grating used for Raman detection into the optical path through the computer-controlled grating tower wheel 15, and turning on the first laser 1 of 633nm;

e, moving the half mirror 27 in the dual-port optical path switching device 10 into the optical path;

f. Observing the sample through the video camera 11, and correcting the detection position in combination with observing the position of the light spot on the sample with the naked eye, and driving the part of the sample that is used for Raman detection by the two-dimensional translation stage 13 to receive laser irradiation;

g, moving the half mirror 27 in the dual-port optical path switching device 10 out of the optical path;

h, the laser light emitted by the first laser 1 sequentially passes through the neutral filter 4, the first mirror 5, the confocal beam expander 6, the first long-pass dichroic mirror 21, and the confocal microscope 8, and then irradiates the sample , the excited Raman light is sent to the monochromator sequentially through the confocal microscope 8, the first long-pass dichroic mirror 21, the first notch filter 20, the mirror 19, and the focusing mirror 18 to obtain a Raman spectrum;

i, adjust the laser power through the neutral filter 4, and adjust the beam diameter through the confocal beam expander 6 until the Raman spectrum with the best intensity is obtained.

(2) Fluorescence spectrum detection

a, placing a planar solid or liquid sample on the two-dimensional displacement stage 13, the sample may also contain a part for fluorescence or Raman spectrum detection;

b, connecting the fluorescence filter group into the optical path by rotating the filter wheel 24;

c, connecting the 375nm second laser 2 into the optical path through the optical switch 3;

d, connect the grating used for fluorescence detection to the optical path by controlling the grating tower wheel 15, and turn on the second laser 2 of 375nm;

e, moving the half mirror 27 in the dual-port optical path switching device 10 into the optical path;

f. Observing the sample through the video camera 11, and correcting the detection position in combination with observing the position of the light spot on the sample with the naked eye, and driving the part of the sample used for fluorescence detection by the two-dimensional displacement stage 13 to receive laser irradiation;

g, moving the half mirror 27 in the dual-port optical path switching device 10 out of the optical path;

h, the laser light emitted by the laser passes through the neutral filter 4, the first mirror 5, the confocal beam expander 6, the second long-pass dichroic mirror 23, and the confocal microscope 8, and then irradiates the sample and is excited. The emitted Raman light is transmitted to the monochromator through the confocal microscope 8, the second long-pass dichroic mirror 23, the second notch filter 22, the reflector 19, and the focusing mirror 18 in order to obtain the fluorescence spectrum;

i, adjust the laser power through the neutral filter 4, and adjust the beam diameter through the confocal beam expander 6 until the fluorescence spectrum with the best intensity is obtained.

The present invention proposes a Raman/fluorescence dual-light-path spectrum detection device. Real Raman detection and fluorescence detection share the same light path. The device realizes the effective combination of fluorescence spectrometer and Raman spectrometer, and is applicable to both Raman and fluorescence Detection, simple structure, and greatly improved space utilization. The function can be switched freely. Fluorescence detection and Raman detection can use the same sample. At the same time, the operator can judge the detection site of the sample through the video camera. The system structure is simple and the cost of use is low. The field has broad application prospects. Neutral filter 4 can adjust the excitation light power to 25%, 50%, 75% and 100% of the initial power, avoiding the central wavelength shift that may be caused by adjusting the optical power through the power adjustment knob that comes with the laser question.

The above are only preferred embodiments of the present invention, and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and changes. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims (10)

1. A Raman/fluorescence dual optical path spectrum detection device is characterized in that it comprises an optical switch (3), a first mirror (5), a confocal microscope (8), a focusing mirror (18), and a third mirror (19), filter structure and monochromator; A sample is provided on the confocal microscope (8); the first laser or the second laser light is irradiated after passing through the optical switch (3), the first mirror (5), the filter structure and the confocal microscope (8) in sequence On the sample, the excited light in the sample passes through the confocal microscope (8), the filter structure, the third mirror (19) and the focusing mirror (18) and then enters the monochromator to obtain the spectrum. 2. Raman/fluorescence dual optical path spectrum detection device according to claim 1, characterized in that a neutral filter is provided between the optical switch (3) and the first reflector (5) (4); A confocal beam expander (6) is provided between the first reflection mirror (5) and the filter structure. 3. Raman/fluorescence dual optical path spectrum detection device according to claim 2, is characterized in that, described filter structure comprises Raman filter set, fluorescence filter set and filter wheel (24) , the Raman filter group and the fluorescence filter group are arranged on the filter wheel (24); the Raman filter group includes the first long-pass dichroic mirror (21 ) and the first notch filter (20), the fluorescence filter group includes the second long-pass dichroic mirror (23) and the second notch filter (22); After the first laser passes through the confocal beam expander (6), it is switched to a Raman filter group by the filter wheel (24), and the first long-pass dichroic mirror (21) reflects the first laser to After the confocal microscope (8), the Raman light with a wavelength greater than the first laser light transmitted by the confocal microscope (8) is received, and the first laser light is filtered out by the first notch filter (20); Or, after the second laser passes through the confocal beam expander (6), it is switched to a fluorescence filter group by the filter wheel (24), and the second long-pass dichroic mirror (23) reflects the second laser After arriving at the confocal microscope (8), it receives fluorescence from the confocal microscope (8) whose wavelength is greater than that of the second laser, and then passes through the second notch filter (22) to filter out the second laser. 4. Raman/fluorescence dual optical path spectrum detection device according to claim 3, is characterized in that, described Raman filter group, described fluorescence filter group, described filter wheel (24) , the outside of the reflector (19) and the focus mirror (18) are packaged in a black interface box (7). 5. Raman/fluorescence dual optical path spectrum detection device according to claim 3, is characterized in that, described confocal microscope (8) comprises second mirror (9), dual-port optical path switching device (10), display A micro-objective lens (12) and a two-dimensional displacement stage (13); the dual-port optical path switching device (10) includes a first displacement mechanism, a first position limiter and a half-mirror (27); the half-mirror (27) and the first position limiter are both arranged on the first displacement structure; the two-dimensional displacement stage (13) is vertically arranged on the plane of the main optical axis of the confocal microscope (8); After the Raman light passes through the second reflector (9), the half-transparent mirror (27) is moved out of the optical path, and the two-dimensional displacement stage (13) drives the sample to receive the Raman light irradiation, and the excited Raman light Sequentially transmit to the monochromator through the first long-pass dichroic mirror (21), the first notch filter (20), the mirror (19) and the focusing mirror (18), to obtain the Raman spectrum; Or, after the fluorescence passes through the second reflection mirror (9), the half-transparent mirror (27) is moved out of the optical path, and the two-dimensional displacement stage (13) drives the sample to receive the fluorescence irradiation, and the excited fluorescence passes through the second mirror in turn. The long-pass dichroic mirror (23), the second notch filter (22), the mirror (19) and the focusing mirror (18) are sent to the monochromator to obtain the fluorescence spectrum. 6. Raman/fluorescence dual optical path spectrum detection device according to claim 5, is characterized in that, a first video camera (11) is provided on one side of the first displacement structure; When the Raman light/fluorescence passes through the second mirror (9), the half-mirror (27) is moved into the optical path, and the signal light enters the video camera (11) after being split by the half-mirror (27). 7. Raman/fluorescence dual optical path spectrum detection device according to claim 5, is characterized in that the first laser is a 633nm laser, which is produced by the first laser (1); The second laser is a 532nm laser, which is generated by the second laser (2), and the second laser emitted by the second laser (2) is reflected to the neutral filter by the fourth reflector (25) under the action of the optical switch (3). in sheet (4). 8. Raman/fluorescence dual optical path spectral detection device according to claim 1, is characterized in that, described monochromator comprises the first concave reflector (16), the second concave reflector (17) and grating tower wheel (15), a plurality of gratings for dispersing light are installed on the grating cone wheel (15); After Raman light or fluorescence passes through the focusing mirror (18) successively through the first concave reflector (16), the grating tower wheel (15) and the second concave reflector (17), the spectrum is obtained; A photoelectric converter (14) is connected to the output end of the second concave reflector (17). 9. Raman/fluorescence dual optical path spectrum detection device according to claim 1, is characterized in that, described optical switch (3) comprises second displacement mechanism, second position limiter and the fifth mirror (26); Both the fifth reflecting mirror (26) and the second position limiter are installed on the second displacement mechanism. 10. A Raman/fluorescence dual-light path spectral detection method, characterized in that the Raman/fluorescence dual-light path spectral detection device described in any one of claims 1 to 9 comprises the following steps: The sample is placed on the confocal microscope (8), and the first laser/second laser is connected to the optical path through the optical switch (3); The first mirror (5), the confocal microscope (8), the focusing mirror (18), the third mirror (19), the filter structure and the monochromator are all connected to the optical path; The first laser/second laser sequentially passes through the first mirror (5), filter structure and confocal microscope (8) and then irradiates the sample, and the excited Raman light passes through the confocal microscope (8), filter After the light structure, the third reflecting mirror (19) and the focusing mirror (18) are passed into the monochromator, the spectrum is obtained.

Images