CN110118726A - A kind of method and apparatus of parallel detecting fluorescent emission difference micro-imaging - Google Patents
A kind of method and apparatus of parallel detecting fluorescent emission difference micro-imaging Download PDFInfo
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- CN110118726A CN110118726A CN201910292539.1A CN201910292539A CN110118726A CN 110118726 A CN110118726 A CN 110118726A CN 201910292539 A CN201910292539 A CN 201910292539A CN 110118726 A CN110118726 A CN 110118726A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N21/64—Fluorescence; Phosphorescence
Abstract
The invention discloses a kind of method and apparatus of parallel detecting fluorescent emission difference micro-imaging, specifically: laser issues laser beam, is converted to line polarisation after being collimated；Phase-modulation is carried out to linearly polarized light, modulation pattern is 0 phase diagram, then is converted into after circularly polarized light to be incident upon on sample to be tested and carries out two-dimensional scanning；The fluorescence signal that the sample to be tested issues is collected using detector array, normalized obtains parallel detecting fluorescence signal light intensity；Modulation pattern is switched to vortex phase figure, phase-modulation is carried out to the linearly polarized light again, repeats the above steps and obtains parallel detecting fluorescence signal light intensity again；Parallel detecting differential signal light intensity is obtained finally, the parallel detecting fluorescence signal light intensity that twice sweep obtains is subtracted each other.High resolution of the invention, signal-to-noise ratio are good, while can be converted by traditional confocal microscope system to very simple, and easy to operate, and the demand to optical power is low.
Description
Technical field
The invention belongs to the micro- field of optical ultra-discrimination, in particular to a kind of parallel detecting fluorescent emission difference micro-imaging
Method and apparatus.
Background technique
Since these centuries, optical microscopy has played important function in science, for people observe microstructure and
Its movement provides a kind of feasible method.However, the microscopical resolution capability of conventional fluorescent is limited in by Abbe diffraction limit
The about half of illumination light wavelength limits the Microstructure observation that length dimension is less than 100nm.For this purpose, people invented it is various
Super-resolution microtechnic, and confocal microscopic imaging technology is the one kind being wherein most widely used.Confocal imaging system height uses
Rate height mainly since the technology can generate the optical section image with high contrast, breaches ordinary optical microscope and spreads out
The limitation of emitter-base bandgap grading limit, lateral resolution is 1.4 times of the ordinary optical microscope of identical numerical aperture, and longitudinal resolution can be with
Reach sub-micrometer scale, while there is simplicity, multi-functional and non-invasive feature, can satisfy various samples and application needs
It asks.
On the basis of confocal microscopic imaging, fluorescence difference micro-imaging technique first uses a solid beam spot scans sample to swash
It fluoresces, collects fluorescence signal, then solid hot spot is modulated to hollow light spot and is used to scan sample excitation fluorescence, second of collection
Two fluorescence signals are finally done difference processing by fluorescence signal, obtain the higher sample image of resolution ratio.
However fluorescence difference micro-imaging technique and confocal microscope are all the light filtered out other than focal plane using pin hole
Signal, thus realize light dicing effect, but the focal plane signal that too small pin hole will lead to collection is too weak, to reduce noise
Than；Biggish pin hole will affect its optical section effect, to limit resolution ratio, therefore people must pass through tradeoff system
Signal-to-noise ratio and resolution ratio determine suitable pinhole size.Also (document Ma Y, Kuang C, Fang Y, et are detailed in horse before
al.Virtual fluorescence emission difference microscopy based on photon
Reassignment [J] Optics Letters, 2015,40 (20): 4627.), Ge Baoliang (be detailed in document Ge B, Wang
Y, Huang Y,et al.Three-dimensional resolution and contrast-enhanced confocal
Microscopy with array detection [J] .Optics Letters, 2016,41 (9): 2013.) and Li Yicheng
(it is detailed in document Li Y, Liu S, Liu D, et al.Image scanning fluorescence emission
difference microscopy based on a detector array[J].Journal of Microscopy,
2017.) the several of proposition are compared based on parallel detecting with the method for difference microtechnic, and the present invention is due to solid spot and hollow spot
All using parallel detecting technology, (horse also illuminates adduction row Detection Techniques using hollow spot, and Ge Baoliang and Li Yicheng are adopted for illumination imaging
Adduction row Detection Techniques are illuminated with solid spot), therefore have higher resolution ratio and signal-to-noise ratio, to can finally obtain matter
Measure higher micro-image.
Summary of the invention
It is an object of the present invention to provide a kind of methods of parallel detecting fluorescent emission difference micro-imaging, can using this method
To be provided simultaneously with the resolution advantage of small pin hole and the collection efficiency of big pin hole, can be improved under the premise of keeping high-resolution
The signal-to-noise ratio of image.
Another object of the present invention is to provide a kind of parallel detecting fluorescent emission difference micro-imaging for realizing the above method
Device, the device can be used for realizing the above method, and fluorescence difference micro-imaging skill is replaced using a fiber-optical probe array
The pin hole and single detector of art, then with a solid beam spot scans sample excitation fluorescence, by spy each in detector array
It surveys the fluorescence signal that device is collected into and carries out image procossing, synthesize the image of a high-resolution and high s/n ratio；Space is used again
Incident solid hot spot is modulated to hollow light spot by optical modulator, is projected and is excited fluorescence on sample, again with same method
The fluorescence signal that detector each in detector array is collected into carries out image procossing, synthesizes second image, finally will
Two images carry out difference processing, to obtain the image of higher resolution.
To achieve the goals above, the method for the fluorescent emission difference micro-imaging of parallel detecting provided by the invention includes
Following steps:
1) laser issues laser beam, is converted to line polarisation after being collimated；
2) phase-modulation is carried out to the linearly polarized light, modulation pattern is 0 phase diagram, modulation function f1(r, θ)=
0, wherein r is the distance that optical axis is arrived in certain point in light beam, and θ cuts open position polar coordinate vectors and pole in plane for the vertical optical axis
The angle of axis；
3) linearly polarized light after the phase-modulation is converted into circularly polarized light using two quarter-wave plates；
4) circularly polarized light, which is incident upon on sample to be tested under the modulation of two-dimensional scanning mirrors system, carries out two-dimensional scanning；
5) fluorescence signal that the sample to be tested issues during two-dimensional scanning is collected using detector array, and will be outer
The signal for enclosing detector collection is moved back to center, then is normalized after all images are added up and obtains parallel detecting fluorescence
Signal light intensity I1(x, y), wherein x, y are the two-dimensional coordinate of scanning element on sample；
6) modulation pattern in step 2) is switched to vortex phase figure, modulation function f1(r, θ)=θ, wherein r
For the distance of certain point in light beam to optical axis, θ is the folder for cuing open position polar coordinate vectors and polar axis in plane of the vertical optical axis
Angle；
7) step 3) is repeated, step 4) collects the sample to be tested using detector array in two-dimensional scanning process again
The fluorescence signal of middle sending, and the signal that peripheral detector is collected is moved back to center, normalizing is carried out after all images are added up
Change processing, obtains parallel detecting fluorescence signal light intensity I again2(x, y), wherein x, y are the two-dimensional coordinate of scanning element on sample；
8) finally, the parallel spy that the parallel detecting differential signal light intensity I (x, y) needed for us can be obtained by twice sweep
It surveys fluorescence signal light intensity to be calculated, calculation formula is I (x, y)=I1(x, y)-β I2(x, y), wherein β be empirical parameter, one
As be set to 0.7；
Wherein, the line polarisation is p-polarization light, because spatial light modulator can only modulate p-polarization light.
Wherein, p polarisation is again converted to rotatory polarization to be scanned to be to make to project the hot spot on sample more evenly.
It wherein, is to keep the degree of polarization of modulated rotatory polarization more preferable using two quarter-wave plates.
Wherein, the scanning range of visual field as needed setting two-dimensional scanning mirrors system.
Wherein, modulated laser beam is Gaussian beam in step 2).
Wherein, modulated laser beam is hollow beam in step 6).
Wherein, when the parallel detecting differential signal light intensity value I (x, y) is negative, it is enabled to be equal to zero.
The principle of the present invention is as follows:
Laser issues laser beam, is converted to line polarisation after being collimated；Phase-modulation is carried out to the linearly polarized light,
Modulation pattern is 0 phase diagram, then is converted into after circularly polarized light to be incident upon on sample to be tested and carries out two-dimensional scanning；Use detection
Device array collects the fluorescence signal that the sample to be tested issues, and the signal that peripheral detector is collected into is moved back to center, then will
All images are normalized after adding up obtains parallel detecting fluorescence signal light intensity；Modulation pattern is switched to vortex phase
Bitmap carries out phase-modulation to the linearly polarized light again, repeats the above steps and obtains parallel detecting fluorescence signal light intensity again；
Parallel detecting differential signal light intensity is obtained finally, the parallel detecting fluorescence signal light intensity that twice sweep obtains is subtracted each other, thus
To the image of higher resolution.
To realize above-mentioned another object, the present invention also provides a kind of dresses of parallel detecting fluorescent emission difference micro-imaging
It sets, including generating the lighting system of excitation beam and collecting the detection system of sample sending fluorescence signal.
On the optical axis of illumination path, it is successively arranged:
For generating the laser of laser beam；
The collimator objective of laser beam collimation for issuing the laser；
For the laser beam after the collimation to be converted to the polarizer of line polarisation；
For the line polarisation to be converted to the half wave plate of p polarisation；
For carrying out the spatial light modulator of phase-modulation to the p polarisation；
For the p polarisation to be converted to the quarter-wave plate of rotatory polarization；
For reflecting the dichroic mirror of the laser beam；
Azimuth and deflection optical path for changing the rotatory polarization, to carry out the scanning galvanometer of two-dimensional scanning to sample
System；
For eliminating the scanning lens of the distortion of the rotatory polarization after being scanned galvanometer system；
Field for collimating and expanding, and being conjugated galvanometer and object lens entrance pupil face the rotatory polarization for being scanned lens
Mirror；
For the rotatory polarization after field lens collimates to be focused to scan the object lens of sample；
For placing the sample stage of sample to be tested.
On the optical axis of detection optical path, it is successively arranged:
For placing the sample stage of sample to be tested；
For collecting the object lens for the fluorescence signal that sample issues on sample stage；
For by the field lens of the fluorescent foci by object lens；
For by the fluorescence collimation and shrink beam by field lens, and the scanning for being conjugated galvanometer and object lens entrance pupil face is saturating
Mirror；
Azimuth and deflection optical path for changing the fluorescence, to solve the scanning galvanometer system of scanning；
For transmiting the dichroic mirror of the fluorescence signal；
For filtering out the optical filter of the stray light of dichroic mirror transmission；
For the fluorescent light beam for passing through optical filter to be focused on to the condenser lens on multimode fiber array；
For acquiring the detector array of the fluorescence signal.
It is additionally provided with the controller for controlling the spatial light modulator and scanning galvanometer system and described for handling
The computer of fluorescence signal；
Sample stage, object lens, field lens, the scanning lens, scanning galvanometer system being arranged on the illumination path and detection optical path
Same device is referred both to dichroic mirror.
The incident light of the spatial light modulator and the angle of emergent light answer it is as small as possible, with reduce due to light pass through it is more than
Crosstalk effect caused by one pixel region and make phase stroke close to design value.
Preferably, the incident light of the spatial light modulator and the angle of emergent light are 5 degree.
0 phase pattern can be switched in the spatial light modulator and vortex phase figure, modulation function are respectively f1(r, θ)=
0 and f1(r, θ)=θ, wherein r is the distance that optical axis is arrived in certain point in light beam, and θ cuts open position pole in plane for the vertical optical axis
The angle of coordinate vector and polar axis；
The multimode fibre plays spatial filter, i.e. pin hole；
Every root multimode fiber in the multimode fiber array is connected with a detector in detector array；
Preferably, the multimode fiber array includes in center multimode fibre and circular row in the center multimode light
The multimode fibre of at least two layers arrangement in a ring outside fibre.In the present invention, the multimode fiber array arrangement mode such as attached drawing 2
Shown, have three layers 19 root multimode fibers altogether；Including a center multimode fibre, arrange in a ring around the center multimode fibre
Two layers of multimode fibre and three layers of multimode fibre, wherein the quantity of two layers of multimode fibre is 6, three layers of multimode fibre are 12.
The detector array is made of avalanche photodide (APD)；
Preferably, the numerical aperture (NA) of the object lens is 1.4；
Preferably, the effective diameter of multimode fiber array is about an Airy disk size；
Preferably, the internal diameter of simple optical fiber is about 0.2 Airy disk size in multimode fiber array.
The present invention compares prior art and has the advantage that
(1) it is provided simultaneously with the resolution advantage of small pin hole and the collection efficiency of big pin hole, can keep high-resolution
Under the premise of improve image signal-to-noise ratio；
(2) it can be improved by traditional confocal microscope system to very simple, and simple to operate；
(3) super resolution image of higher resolution can be obtained with lower optical power.
Detailed description of the invention
Fig. 1 is the schematic device of parallel detecting fluorescent emission difference micro-imaging of the present invention；
Fig. 2 is multimode fiber array schematic device in the present invention；
Fig. 3 is the PSF of the solid laser beam in the present invention；
Fig. 4 is the PSF of the hollow laser beam in the present invention；
Fig. 5 is that parallel detecting differential fluorescence signal light intensity is bent compared with the PSF of confocal signal light intensity normalization in the present invention
Line.
Fig. 6 is parallel detecting differential fluorescence signal light intensity and other several parallel detecting difference microscopic signal light in the present invention
Strong PSF comparison curves.
Specific embodiment
Below with reference to embodiment and attached drawing, the present invention will be described in detail, but the present invention is not limited to this.
A kind of device of the fluorescent emission difference micro-imaging based on parallel detecting, as shown in Figure 1, comprising: laser 1,
Single mode optical fiber 2, collimation lens 3, the polarizer 4, half wave plate 5, the first reflecting mirror 6, spatial light modulator 7, the second reflection
Mirror 8, third reflecting mirror 9, the first quarter-wave plate 10, the second quarter-wave plate 11, dichroic mirror 12, two-dimensional scanning mirrors system
System 13, scanning lens 14, field lens 15, the 4th reflecting mirror 16, object lens 17, sample stage 18, filter plate 19, condenser lens 20, optical fiber
Array 21, detector array 22, computer 23.
The device of the invention embodiment is broadly divided into three parts: generating the lighting system of excitation beam, collects sample sending
The detection system and processor of fluorescence signal, the processor of the present embodiment are computer 23.
Wherein, laser 1, single mode optical fiber 2, collimation lens 3, the polarizer 4, half wave plate 5, the first reflecting mirror 6,
Spatial light modulator 7, the second reflecting mirror 8, third reflecting mirror 9, the first quarter-wave plate 10, the second quarter-wave plate 11,
Dichroic mirror 12, two-dimensional scanning mirrors system 13, scanning lens 14, field lens 15, the 4th reflecting mirror 16, object lens 17, sample stage 18 according to
It is secondary to be arranged on the optical axis of lighting system；
Wherein, sample stage 18, object lens 17, the 4th reflecting mirror 16, field lens 15, scanning lens 14, two-dimensional scanning mirrors system
13, dichroic mirror 12, filter plate 19, condenser lens 20, fiber array 21, detector array 22 is successively set on the light of detection system
On axis；
Wherein, computer 23 is used to control the modulation pattern switching of spatial light modulator 7, two-dimensional scanning mirrors system 13
The signal acquisition of scanning and detector array 22；
It is as follows using the micro- method of parallel detecting fluorescent emission difference using device shown in FIG. 1:
1) laser 1 issues laser beam (it is the feux rouges of 635nm as exciting light that the present embodiment, which uses wavelength) and is coupled
Into single mode optical fiber 2, then lens 3 being collimated after the outgoing of single mode optical fiber 2 and are collimated, the light beam after collimation becomes line by the polarizer 4
Polarised light, linearly polarized light are modulated to p-polarization light by half wave plate 5, are then reflected into space by the first reflecting mirror 6
On optical modulator 7.
2) 7 pairs of spatial light modulator incident p-polarization lights carry out phase-modulation, and modulation pattern is 0 phase diagram, modulation
Function is f1(r, θ)=0, wherein r is the distance that optical axis is arrived in certain point in light beam, θ for the vertical optical axis to cut open plane upper
Set the angle of polar coordinate vectors and polar axis.
3) modulated p-polarization light is reflected into the first quarter-wave plate 10 by the second reflecting mirror 8 and third reflecting mirror 9,
P-polarization light after the phase-modulation is converted to circular polarization by the first quarter-wave plate 10 and the second quarter-wave plate 11
Light, then two-dimensional scanning mirrors system 13 is reflected by dichroic mirror 12.
4) two-dimensional scanning mirrors system 13 changes azimuth and the deflection optical path of incident rotatory polarization, two-dimensional scanning mirrors system
The rotatory polarization of system outgoing, which is scanned after lens 14, eliminates distortion, using field lens 15 collimation and expand, by the 4th reflecting mirror
16 are reflected on object lens 17, inspire fluorescence signal on the sample to be tested being focused on sample stage 18 finally by object lens 17.
5) fluorescence signal of the sample to be tested transmitting on sample stage 18 is collected by object lens 17, anti-by the 4th reflecting mirror 16 later
It is mapped on field lens, reaches scanning galvanometer system 13, quilt after solution scanning using the focusing of field lens 15 and the collimation of scanning lens 14
Dichroic mirror 12 is transmitted through filter plate 19, filters out and is focused lens 20 after stray light and focuses on multimode fiber array 21.Multimode light
Fibre array arrangement mode is as shown in Fig. 2, wherein each roundlet represents a root multimode fiber, and is connected with a detector, directly
Diameter is about 0.2 Airy disk, and the diameter of entire multimode fiber array is about 1 Airy disk.It is collected using detector array 22 glimmering
Light, and the signal that peripheral detector is collected is moved back to center with computer 23, then be normalized after all images are added up
Processing obtains parallel detecting fluorescence signal light intensity I1(x, y), wherein x, y are the two-dimensional coordinate of scanning element on sample.This is visited parallel
Survey fluorescence signal light intensity I1The psf of (x, y) is as shown in Figure 3.
6) modulation pattern in step 2) is switched to vortex phase figure, modulation function f1(r, θ)=θ, wherein r
For the distance of certain point in light beam to optical axis, θ is the folder for cuing open position polar coordinate vectors and polar axis in plane of the vertical optical axis
Angle.
7) step 3), step 4) and step 5) are repeated, obtains parallel detecting fluorescence signal light intensity I again2(x, y), wherein
X, y are the two-dimensional coordinate of scanning element on sample.Parallel detecting fluorescence signal light intensity I2The psf of (x, y) is as shown in Figure 4.
8) finally, the parallel spy that the parallel detecting differential signal light intensity I (x, y) needed for us can be obtained by twice sweep
It surveys fluorescence signal light intensity to be calculated, calculation formula is I (x, y)=I1(x, y)-β I2(x, y), wherein β be empirical parameter, one
As be set to 0.7.
The present invention therewith front method psf distribution curve it is more as shown in Figure 5.Wherein, " confocal " to be shown for standard copolymerization coke
The psf of micro- method；" parallel detecting is confocal " is the psf for being copolymerized burnt microscopic method using parallel detecting, i.e., of the present invention
Parallel detecting fluorescence signal light intensity I1The psf of (x, y)；" difference " is the psf that conventional fluorescent emits difference microscopic method；It is " parallel
Detection difference " is the psf of parallel detecting differential signal light intensity I (x, y) of the present invention.As seen from Figure 5, in the present invention
The psf size of the more other three kinds of methods of the psf size of parallel detecting differential signal light intensity I (x, y) is obviously reduced, therefore this
Invention can be further improved resolution ratio.
The present invention is as shown in Figure 6 compared with the psf distribution curve of several parallel detecting difference methods before.Wherein, " parallel
Detection difference horse " be horse it is also proposed that the psf of parallel detecting differential technique, " parallel detecting difference Pueraria lobota " be Pueraria lobota Bao Liang proposition and
The psf and " parallel detecting difference Lee " of row detection differential technique are psf of the Li Yi at the parallel detecting differential technique of proposition, " simultaneously
Row detection difference " is the psf of the parallel detecting differential signal light intensity I (x, y).As seen from Figure 6, parallel in the present invention
The psf size for detecting differential signal light intensity I (x, y) is small compared with the psf size of other three kinds of parallel detecting difference methods, that is,
With higher resolution ratio.Wherein, although horse it is also proposed that the psf of parallel detecting differential technique be closer to the present invention,
It is due to subtracted in its difference one is that the noise of final result will affect by single small pin hole detection imaging
Than.And the present invention is since solid spot and hollow spot illumination imaging all use parallel detecting technology, two in difference all have
There is the advantages of higher signal-to-noise ratio, final result can have both high-resolution and high s/n ratio.
The foregoing is merely preferable implementation examples of the invention, are not intended to restrict the invention, it is all in spirit of that invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of method of parallel detecting fluorescent emission difference micro-imaging, which comprises the following steps:
1) laser issues laser beam, is converted to linearly polarized light after collimation；
2) phase-modulation is carried out to the linearly polarized light, modulation pattern is 0 phase diagram, modulation function f1(r, θ)=0, wherein
R is that the distance of optical axis, θ is the folder for cuing open position polar coordinate vectors and polar axis in plane of the vertical optical axis for certain point in light beam
Angle；
3) linearly polarized light after phase-modulation is converted to circularly polarized light, and is incident upon on sample to be tested and inspires fluorescence signal；
4) fluorescence signal collected enters detector corresponding with multimode fiber array inner fiber quantity by multimode fiber array
In array, normalized obtains parallel detecting fluorescence signal light intensity I1(x, y), wherein x, y are the two dimension of scanning element on sample
Coordinate；
5) modulation pattern in step 2) is switched to vortex phase figure, modulation function f1(r, θ)=θ, wherein r is light beam
To the distance of optical axis, θ is the angle for cuing open position polar coordinate vectors and polar axis in plane of the vertical optical axis for interior certain point；
6) step 3) and step 4) are repeated, obtains parallel detecting fluorescence signal light intensity I again2(x, y), wherein x, y are to sweep on sample
The two-dimensional coordinate of described point；
7) the parallel detecting fluorescence signal light intensity meter that final parallel detecting differential signal light intensity I (x, y) is obtained by twice sweep
It obtains, calculation formula is I (x, y)=I1(x, y)-β I2(x, y), wherein β is empirical parameter.
2. the method for parallel detecting fluorescent emission difference micro-imaging as described in claim 1, it is characterised in that: the line is inclined
Vibration light is p-polarization light.
3. the method for parallel detecting fluorescent emission difference micro-imaging as described in claim 1, it is characterised in that: the step
2) modulated laser beam is Gaussian beam in.
4. the method for parallel detecting fluorescent emission difference micro-imaging as described in claim 1, it is characterised in that: the step
5) modulated laser beam is hollow beam in.
5. the method for parallel detecting fluorescent emission difference micro-imaging as described in claim 1, it is characterised in that: it is described and
When row detection differential signal light intensity value I (x, y) is negative, I (x, y)=0.
6. a kind of device of parallel detecting fluorescent emission difference micro-imaging, lighting system and collection including generating excitation beam
The detection system of sample sending fluorescence signal, it is characterised in that:
It is successively arranged on the optical axis of the lighting system: laser；Laser beam is converted to the collimation object of the p-polarization light of collimation
Mirror, the polarizer and half wave plate；The spatial light modulator of phase-modulation is carried out to p polarisation；P polarisation is converted into rotatory polarization
Quarter-wave plate；Change rotatory polarization azimuth and deflection optical path to sample carry out two-dimensional scanning two-dimensional scanning mirrors system
System；
On the optical axis of the detection optical path, it is successively arranged: collects the object lens of fluorescence signal；By fluorescent foci to multimode fiber array
On condenser lens and acquisition fluorescence signal detector array；
It is additionally provided with the controller of spatial light modulator and two-dimensional scanning mirrors system and the computer for handling fluorescence signal.
7. the device of parallel detecting fluorescent emission difference micro-imaging as claimed in claim 6, it is characterised in that: the sky
Between the incident light of optical modulator and the angle of emergent light answer it is as small as possible.
8. the device of parallel detecting fluorescent emission difference micro-imaging as claimed in claim 6, it is characterised in that: the sky
Between optical modulator 0 phase pattern and vortex phase figure can be switched；
When for 0 phase pattern, modulation function f1(r, θ)=0, wherein r is that the distance of optical axis, θ is for certain point in light beam
The angle for cuing open position polar coordinate vectors and polar axis in plane of the vertical optical axis；
When for vortex phase figure, modulation function f1(r, θ)=θ, wherein r is distance of the certain point to optical axis in light beam, θ
For the angle for cuing open position polar coordinate vectors and polar axis in plane of the vertical optical axis.
9. the device of parallel detecting fluorescent emission difference micro-imaging as claimed in claim 6, it is characterised in that: described is more
Every root multimode fiber in mode fiber array is connected with a detector in detector array.
10. the device of parallel detecting fluorescent emission difference micro-imaging as claimed in claim 6, it is characterised in that: described
Multimode fiber array includes that at least two layers arrangement in center multimode fibre and circular row outside the center multimode fibre is in ring
The multimode fibre of shape.
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