CN104568872A - Fluorescent micro-spectrum imaging system with optical sectioning strength - Google Patents
Fluorescent micro-spectrum imaging system with optical sectioning strength Download PDFInfo
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Abstract
The invention discloses a fluorescent micro-spectrum imaging system with optical sectioning strength. The fluorescent micro-spectrum imaging system comprises a laser emitter, a beam-expanding light path component, a micro-reflector, a dichroscope, a fluorescent micro-spectrum imaging assembly and a spectrograph, wherein the laser emitter is used for emitting a laser beam to the beam-expanding light path component, and the beam-expanding light path component is used for carrying out speckle suppression to obtain an illuminating beam; the illuminating beam generates a structure light field through the micro-reflector; the structure light field is reflected to the fluorescent micro-spectrum imaging assembly through the dichroscope; the fluorescent micro-spectrum imaging assembly comprises an objective lens, an objective lens drive device and an objective table; the objective lens is used for receiving the structure light field and focusing on a sample on the objective table, and exciting a fluorescent beam; the objective lens drive device drives the objective lens to move along the optical axis, so that the objective lens carries out the optical axis scanning; and the spectrograph analyzes and separates the fluorescent beam to obtain spectrum and image information. By virtue of the fluorescent micro-spectrum imaging system with the optical sectioning strength, a three-dimensional image and corresponding one-dimensional spectrum information of the sample structure can be obtained.
Description
Technical field
The present invention relates to a kind of optical system, particularly relate to a kind of fluorescence microscopy spectrum imaging system with optical chromatography ability.
Background technology
Optical microscope has been just one of important tool of human knowledge's microworld since invention always, and it plays very important effect in the ambits such as clinical medicine, biology, materialogy.But traditional optical microphotograph can only obtain the shape information of object as cell, further can not analyze the structure of object and composition.
Spectral imaging technology is a kind of novel multi-vitamin technology for information acquisition combined with spectral technique by imaging technique., significant otherness can be there is on absorption spectrum in the difference of object structures and composition.Space exploration in macroscopical and earth's surface observe in imaging spectral technology become extensively a kind of and effective detection means.
Along with scientific research and the actual needs detected, spectral imaging technology is applied to the focus that microscopic becomes domestic and international each area research personnel concern gradually, especially the biomedical sector such as pathology, cytology, histology.The microspectrum imaging system that spectral imaging technology and micro-imaging technique are combined together to form, not only can observe the form of object, can also provide in order to differentiation of objects structure and composition difference spectrum information, the two-dimensional signal namely obtaining object can also obtain one dimension spectral information simultaneously.It has higher spectral resolution, optical imagery resolution.
Current major part is based on the fluorescence microscopy spectrum imaging system of traditional fluorescence microscope, it is the wide field imaging mode of a kind of illumination, although a width complete image can be obtained within shorter integral time, but due to the interference by object out of focus part, its image does not have three-dimensional chromatography ability, can only obtain two-dimensional image information and one dimension spectral information.Therefore, how to realize getting rid of the interference of object out of focus part, a kind of fluorescence microscopy spectrum imaging system with optical chromatography ability is newly provided, to obtain three-dimensional image information and one dimension spectral information, meeting and be further analyzed the structure and composition of object, is a current problem in the urgent need to address.
Summary of the invention
Embodiment of the present invention technical matters to be solved is, a kind of fluorescence microscopy spectrum imaging system with optical chromatography ability is provided, three-dimensional structure information and one dimension spectral information can be obtained, thus meet the structure and composition of object is analyzed further.
In order to solve the problems of the technologies described above, embodiments provide a kind of fluorescence microscopy spectrum imaging system with optical chromatography ability, it comprises generating laser, expand optical path component, micro-reflector, dichroic mirror, fluorescent microscopic imaging assembly and spectrometer, described generating laser for by radiating laser beams to described in expand optical path component, illuminating bundle is obtained via described expanding after optical path component carries out speckle suppression, described illuminating bundle produces light field through described micro-reflector, described light field is by described dichroic mirror reflects to described fluorescent microscopic imaging assembly, described fluorescent microscopic imaging assembly comprises object lens, objective lens device and objective table, described object lens accept described light field, and described light field to be focused on sample that described objective table is placed and to inspire fluorescence beam, described objective lens device is for driving described object lens along its axis movement, optical axis scanning is carried out to make described object lens, described fluorescence beam is carried out analyzing and is separated and obtains spectrum and image information by described spectrometer.
Wherein, the described optical path component that expands comprises scatterer whirligig, the first lens and the second lens, described scatterer whirligig obtains described illuminating bundle after described laser beam is carried out speckle suppression, described first lens and the second lens are used for evenly expanding described illuminating bundle, to make described illuminating bundle homogenising.
Wherein, the described fluorescence microscopy spectrum imaging system with optical chromatography ability also comprises the first plane mirror, be located between described second lens and described micro-reflector, after adjusting the incident angle of described illuminating bundle, described illuminating bundle is irradiated on described micro-reflector with certain incident angle.
Wherein, the described fluorescence microscopy spectrum imaging system with optical chromatography ability also comprises post lens, described post lens are located between described micro-reflector and described fluorescent microscopic imaging assembly, in order to described light field is imaged in infinite point, be rectangle shape to make the focal zone of described object lens on the object plane of described sample.
Wherein, described objective lens device comprises one dimension translation stage and object lens adjustment rack, and described object lens adjustment rack is located on described one dimension translation stage, and described object lens adjustment rack is connected with described object lens, in order to adjust the position of described object lens.
Wherein, described spectrometer comprises the second plane mirror, the 3rd lens and entrance slit, described second plane mirror is in order to reflect in described fluorescence beam to described 3rd lens, described 3rd lens in order to focus on described fluorescence beam, to make described fluorescence beam focal imaging on described entrance slit.
Wherein, spectrometer also comprises optical filter, and described optical filter is located between described second plane mirror and described 3rd lens, in order to the laser spectrum in fluorescence beam described in further filtering.
Wherein, described spectrometer also comprises spherical reflector and plane grating, described spherical reflector, for collimating the fluorescence beam through described entrance slit and reflexing to described plane grating, carries out fluorescence spectrum dispersion via described plane grating to described fluorescence beam.
Wherein, described spectrometer also comprises toroidal mirror, and described fluorescence beam carries out, after fluorescence spectrum dispersion, focusing on via described toroidal mirror through described plane grating.
Wherein, described spectrometer also comprises image device, and described fluorescence beam carries out focusing on described image device through described toroidal mirror, carries out the record collection of spectrum and image information via described image device.
Implement the embodiment of the present invention, there is following beneficial effect:
The fluorescence microscopy spectrum imaging system of optical chromatography ability that what the embodiment of the present invention provided have produces one-dimentional structure light field by arranging micro-reflector, dichroic mirror is set simultaneously, object lens, objective lens device and objective table, light field to be reflexed on object lens by dichroic mirror and accepted by object lens, this light field focuses on the sample of objective table by object lens, and inspire fluorescence, one dimension translation is carried out by controlling objective table, obtain the two dimensional image of different visual field, controlling objective lens device drives object lens to move along optical axis direction, object lens are made to carry out optical axis scanning.Fluorescence beam is undertaken analyzing and being separated by spectrometer.Carry out image f iotaeld-of-view switching by controlling objective table, objective lens device carries out objective lens optical axis scanning, and spectrometer carries out fluorescence spectrum dispersion, finally can obtain the 3-D view of sample structure and the one dimension spectral information of correspondence.
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, below the accompanying drawing used required in describing embodiment is briefly described, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skill in the art, under the prerequisite not paying creative work, other accompanying drawing can also be obtained according to these accompanying drawings.
Fig. 1 is the schematic diagram with the fluorescence microscopy spectrum imaging system of optical chromatography ability that the embodiment of the present invention provides;
Fig. 2 is the schematic diagram at another visual angle of the post lens that provide of the embodiment of the present invention and micro-reflector;
Fig. 3 is the schematic diagram of the micro-reflector generation light field that the embodiment of the present invention provides;
Fig. 4 is the imaging schematic diagram with the fluorescence microscopy spectrum imaging system of optical chromatography ability that the embodiment of the present invention provides;
Fig. 5 is the timing sequence diagram of micro-reflector, image device and the objective table that the embodiment of the present invention provides.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, be clearly and completely described the technical scheme in the embodiment of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
See also shown in Fig. 1 to Fig. 2, the present embodiment provides a kind of fluorescence microscopy spectrum imaging system 100 with optical chromatography ability, and it comprises generating laser 10, expands optical path component 20, micro-reflector 30, dichroic mirror 40, fluorescent microscopic imaging assembly 50 and spectrometer 60.Described fluorescence microimaging systems 50 comprises object lens 51, objective table 52 and objective lens device 53, described objective table 52 is placed with sample (not shown), described objective lens device 53 drives described object lens 51 along its axis movement, carries out optical axis scanning to make described object lens 51.
Described generating laser 10 is for Emission Lasers bundle L1.In the present embodiment, described generating laser 10 is mono-colour laser.Described generating laser 10 expands in optical path component 20 described in being emitted to by described laser beam L1.The optical maser wavelength of described generating laser 10 is 445nm, and laser power is 40mw.Be understandable that, in other embodiments, the optical maser wavelength of described generating laser 10 and laser power also can select adjustment according to practical operation.
Described expanding after the described laser beam L1 that described generating laser 10 is launched is carried out speckle suppression by optical path component 20 obtains illuminating bundle L2.In the present embodiment, described in expand optical path component 20 and comprise scatterer whirligig 21, first lens 22 and the second lens 23, described scatterer whirligig 21 obtains described illuminating bundle L2 after described laser beam L1 is carried out speckle suppression.Described scatterer whirligig 21 is made up of diffusion sheet (not shown) and electro-motor (not shown), diffusion sheet described in described electrical motor driven rotates, thus by the rotation of described diffusion sheet, upset the laser correlativity of described laser beam L1, thus realize speckle inhibit feature.
Described illuminating bundle L2 evenly expands through described first lens 22 and described second lens 23, to make described illuminating bundle L2 homogenising.In addition, described first lens 22 and the second lens 23 also can collimate described illuminating bundle L2, are convenient to follow-up described micro-reflector 30 and form light field.
Described illuminating bundle L2 produces light field L3 through described micro-reflector 30.In the present embodiment, described micro-reflector 30 is digital micro-mirror, thus can produce described light field L3 fast, and due to the refresh rate of its described light field L3 produced fast, therefore its efficiency of light energy utilization is high.In addition, the described light field L3 produced by described micro-reflector 30 evenly, thus be conducive to follow-up sample structure analysis.Concrete, as shown in Figure 3, described micro-reflector 30 adopts DMD optical projection system, be made up of 1024 × 768 pixels, its pixel dimension is 13.68um × 13.68um, and namely pixel described in each is one piece of small reflector that independently can overturn 12 degree to positive negative direction, and frequency of its upset can reach 32KHz.Each cycle Z of described micro-reflector is 6 pixels.Be understandable that, in other embodiments, described micro-reflector 30 also can select other optical projection system according to practical operation.
In order to further improvement, the described fluorescence microscopy spectrum imaging system 100 with optical chromatography ability also comprises the first plane mirror 70, described first plane mirror 70 is located between described second lens 23 and described micro-reflector 30, after adjusting the incident angle of described illuminating bundle L2, described illuminating bundle L2 is located on described micro-reflector 30 with certain incident angle.In the present embodiment, described incident angle is 12 degree, thus when making described illuminating bundle L2 on described micro-reflector 30, can be modulated to the one-dimentional structure light field with some cycles.Be understandable that, in other embodiments, the value of described incident angle also can adjust according to practical operation.
In the present embodiment, the described fluorescence microscopy spectrum imaging system 100 with optical chromatography ability also comprises post lens 80, described post lens 80 are located between described micro-reflector 30 and described fluorescent microscopic imaging assembly 50, in order to described light field L3 is imaged in infinite point, be rectangle shape to make the focal zone of described object lens 51 on the object plane of described sample.
Described dichroic mirror 40 is located between described post lens 80 and described object lens 51, in order to reflect described light field L3.In the present embodiment, described dichroic mirror 40 can reflect the spectrum being less than 450nm, simultaneously transmission can be greater than the spectrum of 450nm, thus described light field L3 is reflexed on described object lens 51 through described dichroic mirror 40, the described fluorescence beam L4 excited is transmitted through in described spectrometer 60 through described dichroic mirror 40.
Described object lens 51 accept described light field L3, and described light field L3 focuses on the described sample of placement on described objective table 52 by described object lens 51, and inspire fluorescence beam L4.Described objective lens device 53, for driving described object lens 51 along its axis movement, carries out optical axis scanning to make described object lens 51.In the present embodiment, described objective lens device 53 can do quasi-nano along its optical axis direction and not scan.Described objective lens device 53 comprises one dimension translation stage (not shown) and object lens adjustment rack (not shown), described object lens adjustment rack is located on described one dimension translation stage, described object lens adjustment rack is connected with described object lens 51, in order to adjust the position of described object lens 51, thus the positioning precision of described object lens 51 can be ensured.Concrete, the range of described one dimension translation stage is 25mm, and its repetitive positioning accuracy is 1.6um, and minimum step displacement is 50nm.Be understandable that, in other embodiments, the range of described one dimension translation stage and its repetitive positioning accuracy all can be selected according to the adjustment of practical operation situation.
In the present embodiment, the displacement range of described objective table 52 is 110mm × 75mm, and its repetitive positioning accuracy is 0.25um, and described objective table 52 is for scanning described sample image thus obtaining the two dimensional image of whole sample.Be understandable that, in other embodiments, the displacement range of described objective table 52 and its repetitive positioning accuracy all can be selected according to the adjustment of practical operation situation.
In the present embodiment, when described object lens 51 accept described light field L3 and focus on the sample of described objective table 52, described light field L3 inspires fluorescence beam L4, described fluorescence beam L4 and laser are again through described dichroic mirror 40, described laser is reflected by described dichroic mirror 40 again, described fluorescence beam L4 is then incident on described spectrometer 60 through described dichroic mirror 40, to carry out analyzing and being separated.
Described fluorescence beam L4 carries out analyzing and is separated and obtains spectrum and image information by described spectrometer 60.In the present embodiment, the spectral range of described spectrometer 60 is 400 ~ 700nm, and grating line number is 600lp/mm, and spectral resolution is better than 2nm.Be understandable that, in other embodiments, the spectral range of described spectrometer 60, grating line number and spectral resolution all can be selected according to practical operation adjustment.
Described spectrometer 60 comprises the second plane mirror 61, the 3rd lens 62 and entrance slit 63, described second plane mirror 61 is in order to reflect on described fluorescence beam L4 to described 3rd lens 62, described 3rd lens 62 in order to focus on described fluorescence beam L4, to make described fluorescence beam L4 focal imaging on described entrance slit 63.In the present embodiment, the image planes at described micro-reflector 30 place plane, described sample place object plane, described entrance slit 63 place plane and image device 19 place, four one-tenth mutual conjugate relations.Meanwhile, there is Linear Amplifer relation in the object plane visual field size of described sample and described entrance slit 63.
In the present embodiment, described spectrometer 60 also comprises optical filter 64, and described optical filter 64 is located between described second plane mirror 61 and described 3rd lens 62, in order to the remaining laser spectrum in fluorescence beam L4 described in further filtering.
In the present embodiment, described spectrometer 60 also comprises spherical reflector 65 and plane grating 66, described spherical reflector 65, in order to collimate the fluorescence beam L4 through described entrance slit 63 and to reflex on described plane grating 66, carries out fluorescence spectrum dispersion via described plane grating 66 to described fluorescence beam L4.Carry out spectral dispersion by described plane grating 66, thus the wavelength coverage of spectrum is longer, spectral accuracy is higher, and the linear relationship of spectrum and pixel is better.
In order to further improvement, in the present embodiment, described spectrometer 60 also comprises toroidal mirror 67, and described fluorescence beam L4, after described plane grating 66 carries out fluorescence spectrum dispersion, focuses on via described toroidal mirror 67.In addition, described toroidal mirror 67 can carry out the correction of astigmatism and coma to the described fluorescence microscopy spectrum imaging system 100 with optical chromatography ability, thus has the imaging performance of the fluorescence microscopy spectrum imaging system 100 of optical chromatography ability described in improving.
In the present embodiment, spectrum and image information is gathered for the ease of record, described spectrometer 60 also comprises image device 68, and described fluorescence beam L4 focuses on described image device 68 through described toroidal mirror 67, carries out the record collection of spectrum and image information via described image device 68.
The fluorescence microscopy spectrum imaging system 100 with optical chromatography ability provided by the invention produces light field L3 by arranging micro-reflector 30, and dichroic mirror 40 is set the light field L3 that described micro-reflector 30 produces is reflexed on object lens 51, the described light field L3 received focuses on the sample of described objective table 52 by described object lens 51.Carry out one dimension translation by controlling described objective table 52, thus obtain the two dimensional image of different visual field, control described objective lens device 53 and drive described object lens 51 to move along optical axis direction, thus make described object lens 51 to carry out optical axis scanning.
Referring to Fig. 4, is the imaging schematic diagram with the fluorescence microscopy spectrum imaging system 100 of optical chromatography ability of the present invention, described in there is the fluorescence microscopy spectrum imaging system 100 of optical chromatography ability mode of operation be optical chromatography pattern.During work, expand in optical path component 20 described in laser beam L1 is emitted to by described generating laser 10, speckle suppression is carried out and after evenly expanding via the described optical path component 20 that expands, form illuminating bundle L2, described illuminating bundle L 2 is radiated on described micro-reflector 30, thus produce light field L3, and this light field L3 is the one-dimentional structure light field with some cycles.After described light field L3 images in infinite distance via described post lens 80, to reflex on described object lens 51 through described dichroic mirror 40 and accepted by described object lens 51, described light field L3 focuses on the sample of described objective table 52 by described object lens 51, and inspiring fluorescence beam L4, the illuminated scope of the object plane on described objective table 52 is one piece of rectangular field of view.
Described fluorescence beam L4 reflexes on described 3rd lens 62 by described second plane mirror 61, and after focusing on the imaging of described entrance slit 63, by spherical mirror 65 collimated reflected on plane grating 66, spectral dispersion is carried out by described plane grating 66, and focus to record and the collection of described image device 68 carrying out image information and spectrum via described toroidal mirror 67, thus obtain the monochromatic tomographic map of the different spectrum of current place optical axis scanning slice.。
See also Fig. 3 to Fig. 5, in the present embodiment, when the described mode of operation with the fluorescence microscopy spectrum imaging system 100 of optical chromatography ability is optical chromatography pattern, described in have the operation steps of the fluorescence microscopy spectrum imaging system 100 of optical chromatography ability as follows:
(1) control the light field that described micro-reflector 30 order loads three width outs of phase, and make light field distribution arrangement consistent with the Width of described object plane rectangle visual field, as shown in Figure 3.
(2) adopt CCD camera (not shown) to gather the two-dimension spectrum figure of the described light field of three corresponding width outs of phase simultaneously
and
(3) within the time that described spectrogram reads by described CCD camera for the third time, control objective table 52 simultaneously and carry out one dimension translation, and switching focusing is in the sample image of described entrance slit 63, namely switches next visual field and enters described entrance slit 63.
(4) timing sequence diagram of described micro-reflector 30, image-forming assembly 68 and objective table 52 as shown in Figure 5.Repeat above process, the two-dimension spectrum image of one group of different visual field can be obtained, by can obtain the monochrome two-dimensional image group of different spectrum to the described two-dimension spectrum image reconstruction of the different visual fields gathered, recycle the monochromatic tomographic map that can obtain the different spectrum of sample after following formula carries out image operation.
Wherein, I
rErepresent be rebuild after spectrum.
(5) controlling described objective lens device 53 drives described object lens 51 to move along its optical axis direction, and repeats the step of above-mentioned (1) ~ (4), thus can obtain the monochromatic tomographic map of the different spectrum of Current Scan layer sample.
(6) repeat the step of above-mentioned (1) ~ (5), the monochromatic tomographic map of different spectrum corresponding to different layers sample can be obtained, be i.e. four-dimensional image information group.
The fluorescence microscopy spectrum imaging system of optical chromatography ability that what the embodiment of the present invention provided have produces one-dimentional structure light field by arranging micro-reflector, dichroic mirror is set simultaneously, object lens, objective lens device and objective table, light field to be reflexed on object lens by dichroic mirror and accepted by object lens, this light field focuses on the sample of objective table by object lens, and inspire fluorescence, one dimension translation is carried out by controlling objective table, obtain the two dimensional image of different visual field, controlling objective lens device drives object lens to move along optical axis direction, object lens are made to carry out optical axis scanning.Fluorescence beam is undertaken analyzing and being separated by spectrometer.Carry out image f iotaeld-of-view switching by controlling objective table, objective lens device carries out objective lens optical axis scanning, and spectrometer carries out fluorescence spectrum dispersion, finally can obtain the 3-D view of sample structure and the one dimension spectral information of correspondence.
The above is the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications are also considered as protection scope of the present invention.
Claims (10)
1. one kind has the fluorescence microscopy spectrum imaging system of optical chromatography ability, it is characterized in that: it comprises generating laser, expand optical path component, micro-reflector, dichroic mirror, fluorescent microscopic imaging assembly and spectrometer, described generating laser for by radiating laser beams to described in expand optical path component, illuminating bundle is obtained via described expanding after optical path component carries out speckle suppression, described illuminating bundle produces light field through described micro-reflector, described light field is by described dichroic mirror reflects to described fluorescent microscopic imaging assembly, described fluorescent microscopic imaging assembly comprises object lens, objective lens device and objective table, described object lens accept described light field, and described light field to be focused on sample that described objective table is placed and to inspire fluorescence beam, described objective lens device is for driving described object lens along its axis movement, optical axis scanning is carried out to make described object lens, described fluorescence beam is carried out analyzing and is separated and obtains spectrum and image information by described spectrometer.
2. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 1, it is characterized in that: described in expand optical path component and comprise scatterer whirligig, the first lens and the second lens, described scatterer whirligig obtains described illuminating bundle after described laser beam is carried out speckle suppression, described first lens and the second lens are used for evenly expanding described illuminating bundle, to make described illuminating bundle homogenising.
3. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 2, it is characterized in that: described in there is optical chromatography ability fluorescence microscopy spectrum imaging system also comprise the first plane mirror, be located between described second lens and described micro-reflector, after adjusting the incident angle of described illuminating bundle, described illuminating bundle is irradiated on described micro-reflector with certain incident angle.
4. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 1, it is characterized in that: described in there is optical chromatography ability fluorescence microscopy spectrum imaging system also comprise post lens, described post lens are located between described micro-reflector and described dichroic mirror, in order to described light field is imaged in infinite point, be rectangle shape to make the focal zone of described object lens on the object plane of described sample.
5. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 1, it is characterized in that: described objective lens device comprises one dimension translation stage and object lens adjustment rack, described object lens adjustment rack is located on described one dimension translation stage, described object lens adjustment rack is connected with described object lens, in order to adjust the position of described object lens.
6. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 1, it is characterized in that: described spectrometer comprises the second plane mirror, the 3rd lens and entrance slit, described second plane mirror is in order to reflect in described fluorescence beam to described 3rd lens, described 3rd lens in order to focus on described fluorescence beam, to make described fluorescence beam focal imaging on described entrance slit.
7. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 6, it is characterized in that: spectrometer also comprises optical filter, described optical filter is located between described second plane mirror and described 3rd lens, in order to the laser spectrum in fluorescence beam described in further filtering.
8. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 7, it is characterized in that: described spectrometer also comprises spherical reflector and plane grating, described spherical reflector, for collimating the fluorescence beam through described entrance slit and reflexing to described plane grating, carries out fluorescence spectrum dispersion via described plane grating to described fluorescence beam.
9. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 8, it is characterized in that: described spectrometer also comprises toroidal mirror, described fluorescence beam carries out, after fluorescence spectrum dispersion, focusing on via described toroidal mirror through described plane grating.
10. there is the fluorescence microscopy spectrum imaging system of optical chromatography ability as claimed in claim 9, it is characterized in that: described spectrometer also comprises image device, described fluorescence beam focuses on described image device through described toroidal mirror, carries out the record collection of spectrum and image information via described image device.
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