CN103940787B - The imaging of dual wavelength Quantization phase and fluorescence imaging association system - Google Patents
The imaging of dual wavelength Quantization phase and fluorescence imaging association system Download PDFInfo
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- CN103940787B CN103940787B CN201310016989.0A CN201310016989A CN103940787B CN 103940787 B CN103940787 B CN 103940787B CN 201310016989 A CN201310016989 A CN 201310016989A CN 103940787 B CN103940787 B CN 103940787B
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Abstract
The invention provides the imaging of a kind of dual wavelength Quantization phase and fluorescence imaging association system.In the present invention, common focusing principle is applied in Quantization phase imaging, fluorescence imaging is combined with Quantization phase imaging, adopt the method for Spatial transmission and dual wavelength modulation, while the signal to noise ratio (S/N ratio) that improve Quantization phase microscopic system and resolution, obtain high-resolution fluoroscopic image, in cell fluorescence imaging, structure imaging, internal structure of body, Surface testing etc., there is good application.
Description
Technical field
The present invention relates to optical field, particularly relate to a kind of novel dual wavelength Quantization phase imaging based on common focusing principle with ultrahigh resolution and fluorescence imaging association system.
Background technology
Modern biomedical research needs the accurate location of observing protein in cell three-dimensional space and distribution.Protein only navigates to its biological function of ability display on special subcellular organelle and physiologically active site, and its location is a dynamic process.Correctly to be familiar with and just must to mark it with the function of Study on Protein thus to study its location change under different physiological status.For reaching this purpose, people first carry out fluorescence labeling to studied molecule usually, and obtain fluorescence imaging, cell transmission imaging of reentrying, utilizes image processing techniques, fluoroscopic image and transmission image are merged, and obtain the location of molecule in cell and function information.Due to the diffraction of optical system, image resolution ratio is not high.
Transmission micro-imaging can provide cellular morphology and structural information, and fluorescent microscopic imaging can provide the function of studied molecule, location and reaction kinetics information.Phase imaging technology utilizes different material refractive index different, the laser of transmission is made to produce phase differential and interfere, by obtaining the refractive index of diverse location in light path to the analysis of interference pattern, and be converted into image, make the resolution of the microscopical optical propagation direction of transmission imaging (Z-direction) that nm magnitude can be reached.Fluorescent microscope is along with the development that laser co-focusing, photosensitive localize fluorescent are micro-, single molecular fluorescence interferes the technology such as fluorescence microscopy, and molecule positioning precision also reaches nm magnitude.
Therefore, further raising phase imaging technology is in the resolution of X-Y plane, and this technology and high-resolution Imaging-PAM are combined, can observe easily and the expression of quantitative objective protein in living cells, location, transhipment and the interaction with other biological molecule, for life science provides new research means.By the imaging of integrated application confocal fluorescent, the positioning precision of photosensitive localize fluorescent imaging technique, Quantization phase imaging microscope is in the precision of cellular prion protein imaging, propose under the prerequisite of common focusing principle, by the concept that Quantization phase imaging and fluorescence imaging are combined, for super-resolution provides scheme in the application of bion, in addition compared with the imaging of Single wavelength Quantization phase, dual wavelength is by carrying out speckle noise balance to image in polyenergetic territory and smoothing processing improves resolution, improve the sensitivity of measuring method, absolute information is extracted by whole audience observation, the different information of object under test are provided by different wave length simultaneously, the problem of the signal ambiguity that unavoidable phase shift brings in Single wavelength record phase imaging can also be solved, the system that substantially increases is at x, y, the resolution of z.
But realizing in process of the present invention, applicant finds, current microscope is difficult to obtain the accurate location of target molecule in cell, namely fluoroscopic image and eucaryotic cell structure image co-registration, cellular structures and functions imaging cannot be realized, thus causes the inconvenience of research work.
Summary of the invention
(1) technical matters that will solve
For solving above-mentioned one or more problems, the invention provides the imaging of a kind of dual wavelength Quantization phase and fluorescence imaging association system.
(2) technical scheme
According to an aspect of the present invention, the imaging of a kind of dual wavelength Quantization phase and fluorescence imaging association system is provided.This association system comprises: light-source system 100, for providing the first laser beam and second laser beam of different wave length; Point tread assembly 200, for being the first reference beam and the first object light light beam by the first laser beam beam splitting; Be the second reference beam and the second object light light beam by the second laser beam beam splitting, the first reference beam and the second reference beam inject reference path 400; First object light light beam and the second object light light beam inject object light light path 300; Object light light path 300, for by the first object light light beam and the second object light beam projection to sample, the first object light light beam and the second object light light beam that carry sample message reflex to interference imaging system 500 by the 5th Amici prism 314; Meanwhile, the fluorescent light beam produced by the first object light light beam and the second object light beam excitation sample is transmitted through fluoroscopic imaging systems 600 through the 6th Amici prism 316; Reference path 400, for being projected to interference imaging system 500 by the first reference beam and the second reference beam; Interference imaging system 500, for by the first reference beam and the first object light light beam, and the second reference beam and the second object light light beam carry out interference imaging; Fluoroscopic imaging systems 600, shares co-focusing imaging part with described object light light path 300, for carrying out imaging to the fluorescent light beam of the first object light light beam and the second object light beam excitation.
(3) beneficial effect
As can be seen from technique scheme, multi electron beam differential frequency multiplication pipe of the present invention has following beneficial effect:
(1) combination of fluorescence imaging and Quantization phase imaging, solves the shortcoming that cellular morphology and structure can not be measured simultaneously, and part light path shares;
(2) adopt dual wavelength light to carry out interference imaging, and carry out record simultaneously with a CCD, overcome the shortcoming that multiple CCD records;
(3) introduce Spatial transmission structure, achieve the super-resolution imaging in x, y, z plane, and achieve the separation of different wave length information;
(4) Confocal laser endomicroscopy principle is applied in Quantization phase imaging, realizes the raising of x, y, z directional resolution.
Accompanying drawing explanation
Fig. 1 is based on the dual wavelength Quantization phase imaging of common focusing principle and fluorescence imaging association system schematic diagram;
Fig. 2 microlens array schematic diagram;
Fig. 3 a is the schematic diagram of the embodiment of the present invention as the rectangular pyramid of laser interference element;
Fig. 3 b adopts the schematic diagram of the laser interference pattern that rectangular pyramid produces as laser interference element shown in Fig. 3 a for the embodiment of the present invention;
Fig. 4 is the embodiment of the present invention laser interference element when being rectangular pyramid, produce the optical axis longitudinal sectional drawing of interference field;
Fig. 5 a is the schematic diagram of the embodiment of the present invention as the truncated rectangular pyramids of laser interference element;
Fig. 5 b adopts the schematic diagram of the laser interference pattern that rectangular pyramid produces as laser interference element shown in Fig. 5 a for the embodiment of the present invention;
[main element symbol description]
100-light-source system; 200-divides tread assembly;
300-object light light path; 400-reference path;
500 interference imaging systems; 600-fluoroscopic imaging systems;
110-first laser instrument; 111-first beam expander;
112-first half-wave plate; 120-second laser;
121-second beam expander; 122-second half-wave plate;
211-first polarization splitting prism; 221-second polarization splitting prism;
311-second catoptron; 312-the 4th Amici prism;
313-microlens array (terrace with edge or pyramid); 314-the 5th Amici prism;
315-pinhole array; 316-the 6th Amici prism;
317-microcobjective; 318-displacement platform;
410-the 4th half-wave plate; 411-second rectangular parallelepiped glass lens;
412-second Quantization phase adjuster; 413-first catoptron;
420-the 3rd half-wave plate; 421-first rectangular parallelepiped glass lens;
422-first Quantization phase adjuster; 423-the 3rd Amici prism;
510-imaging lens group; 511-diaphragm;
512-interference imaging CCD; 610-optical filter;
611-fluorescence imaging lens combination; 612-diaphragm;
613-EMCCD。
Embodiment
For making the object, technical solutions and advantages of the present invention clearly understand, below in conjunction with specific embodiment, and with reference to accompanying drawing, the present invention is described in more detail.
It should be noted that, in accompanying drawing or instructions describe, similar or identical part all uses identical figure number.The implementation not illustrating in accompanying drawing or describe is form known to a person of ordinary skill in the art in art.In addition, although herein can providing package containing the demonstration of the parameter of particular value, should be appreciated that, parameter without the need to definitely equaling corresponding value, but can be similar to corresponding value in acceptable error margin or design constraint.In addition, the direction term mentioned in following examples, such as " on ", D score, "front", "rear", "left", "right" etc., be only the direction with reference to accompanying drawing.Therefore, the direction term of use is used to illustrate and is not used for limiting the present invention.
The invention provides the novel dual wavelength Quantization phase imaging based on common focusing principle with ultrahigh resolution and fluorescence imaging association system, common focusing principle is applied in Quantization phase imaging by this system, and fluorescence imaging is combined with Quantization phase imaging, the advantage of both utilizations, in cell fluorescence imaging, structure imaging, internal structure of body, Surface testing etc., there is good application, for cellular morphology and in site measurement provide a kind of new means.
Details are as follows by reference to the accompanying drawings for a preferred embodiments of the present invention: if Fig. 1 is based on the dual wavelength Quantization phase imaging of common focusing principle and fluorescence imaging association system, comprise light-source system 100, point tread assembly 200, object light light path 300, reference light light path 400, interference imaging system 500 and fluoroscopic imaging systems 600, wherein object light light path 300 and fluoroscopic imaging systems 600 share co-focusing imaging part.Below each assembly is described in detail.
Light-source system 100
Light-source system 100 provides the first laser beam and second laser beam of different wave length, comprising: the first laser instrument 110, first beam expander 111, first half-wave plate 112, second laser 120, second beam expander 121, second half-wave plate 122.
Wherein, first laser instrument 110 sends the first laser beam that wavelength is 346nm, 495nm, 514nm, 556nm, 647nm or 710nm, this first laser beam carries out beam-expanding collimation through the first beam expander 111, then enters beam splitting assembly 200 after the first half-wave plate 112, as shown in phantom in Figure 1.
Wherein, second laser 120 sends the second laser beam (with the first laser beam different wave length) that wavelength is 346nm, 495nm, 514nm, 556nm, 647nm or 710nm, this second laser beam carries out beam-expanding collimation through the second beam expander 121, beam splitting assembly 200 is entered again, as shown in fig. 1 by dash-dotted lines after the second half-wave plate 122.
It will be apparent to those skilled in the art that the wavelength of this first laser beam and the second laser beam can adjust according to the needs of fluorescence excitation.
Divide tread assembly 200
First laser beam and the second laser beam beam splitting are reference beam and object light light beam by point tread assembly 200 respectively, comprise the first polarization splitting prism 211 and the second polarization splitting prism 221 mutually staggered;
First laser beam is the first reference beam and the first object light light beam by the first polarization splitting prism 211 light splitting.Wherein, the first reference beam injects reference path 400; First object light light beam injects object light light path 300.
Second laser beam is the second reference beam and the second object light light beam by the second polarization splitting prism 221 light splitting.Wherein, the second reference beam injects reference path 400; Second object light light beam injects object light light path 300.
Object light light path 300
Object light light path 300 by the first object light light beam and the second object light beam projection on sample, carry the first object light light beam of sample message and the second object light light beam through sample stage 318 reflect simultaneously the 5th Amici prism 314 catoptric imaging to interference imaging system 500.This object light light path comprises the second catoptron 311, the 4th Amici prism 312, microlens array (terrace with edge or pyramid) (as Fig. 2) 313, the 5th Amici prism 314, pinhole array 315, the 6th Amici prism 316, microcobjective 317 and displacement platform 318 and forms.
First object light light beam is after the second catoptron 311 reflects, through the 4th Amici prism 312 transmission, be radiated at microlens array (terrace with edge or pyramid) 313, thus generation has the array of certain space periodic arrangement (as Fig. 3, 5), this array is mated with pinhole array 315 by the 5th Amici prism 314, be radiated on the sample be positioned on displacement platform 318 through the 6th Amici prism 316 and microcobjective 317, thus become the first object light light beam carrying sample message, reflect through displacement platform 318 and carry out imaging by microcobjective 450 again, through the 6th Amici prism 316, elimination veiling glare is being mated with pinhole array 315, finally be projected to the 5th Amici prism 314, through the 5th Amici prism 314 catoptric imaging to interference imaging system 500.
Second object light light beam is after the 4th Amici prism 312 reflects, be radiated at microlens array (terrace with edge or pyramid) 313, thus generation has the array of certain space periodic arrangement (as Fig. 3, 5), this array is mated with pinhole array 315 by the 5th Amici prism 314, be radiated on the sample be positioned on displacement platform 318 through the 6th Amici prism 316 and microcobjective 317, thus become the first object light light beam carrying sample message, reflect through displacement platform 318 and carry out imaging by microcobjective 450 again, through the 6th Amici prism 316, elimination veiling glare is being mated with pinhole array 315, finally be projected to the 5th Amici prism 314, through the 5th Amici prism 314 catoptric imaging to interference imaging system 500.
Wherein the first reference beam and the second reference beam are transmitted to interference imaging system 500 by the 5th Amici prism 314, will carry the first object light light beam of sample message and the second object light beam reflection to interference imaging system 500.
Reference path 400
Reference path 400 regulates the phase place of the first reference beam and the second reference beam respectively, and both are injected the 5th Amici prism 314, through the 5th Amici prism 314 transmission imaging to interference imaging system 500, as shown in the solid line of Fig. 1 the 3rd Amici prism 415 light path rear end.This reference beam system 400 comprises: the 4th half-wave plate 410, second rectangular parallelepiped glass lens 411, second Quantization phase adjuster 412 and the first catoptron 413 of the second polarization splitting prism 221 light path rear end; And be positioned at the 3rd half-wave plate 420, first rectangular parallelepiped glass lens 421, first Quantization phase adjuster 422 and the 3rd Amici prism 423 of the first polarization splitting prism 211 light path rear end.
First reference beam, after the 3rd half-wave plate 420 that is positioned at the first polarization splitting prism 211 rear end and the first rectangular parallelepiped glass lens 421, adjusts phase place by the first Quantization phase adjuster 422, and reflexes to interference imaging system 500 through the 3rd Amici prism 423.
Second reference beam is after the 4th half-wave plate 410 that is positioned at the second polarization splitting prism 221 rear end and the second rectangular parallelepiped glass lens 411, phase place is adjusted by the second Quantization phase adjuster 412, reflect through catoptron 413, and be transmitted through interference imaging system 500 by the 3rd Amici prism 423.
Wherein, quantization phase modulation is the quantization phase modulation of double mirror type, and it moves up and down the light path adjusting the first reference beam and the second reference beam by entirety, and then realizes object light and the small optical path difference of reference light.
Interference imaging system 500
Interference imaging system 500 is by the first reference beam and the first object light light beam, and the second reference beam and the second object light light beam carry out interference imaging, and is received by same CCD.This interference imaging system 500 comprises: imaging lens group 510, diaphragm 511 and interference imaging CCD512 form.
First reference beam with after sample, carry the first object light light beam of sample message through imaging lens group 510 imaging, after diaphragm 511, interference imaging CCD512 interferes, forms the interferogram with object information, as shown in Figure 3.
Equally, the second reference beam with after sample, carry the second object light light beam of sample message through imaging lens group 510 imaging, after diaphragm 511, interference imaging CCD512 interferes, forms the interferogram with object information.
Fluoroscopic imaging systems 600
Fluoroscopic imaging systems 600 can be the fluorescence imaging such as photosensitive localize fluorescent system or stimulated emission depletion micro imaging system, for fluorescent light beam is carried out imaging, comprising: microlens array (terrace with edge or pyramid) the 313, the 5th Amici prism 314, pinhole array 315, the 6th Amici prism 316, microcobjective 317 and displacement platform 318, optical filter 610, fluorescence imaging lens combination 611, diaphragm 612 and EMCCD613.
First object light light beam and the second object light light beam irradiation are at microlens array (terrace with edge or pyramid) 313, thus generation has the array of certain space periodic arrangement (as Fig. 3, 5), this array is mated with pinhole array 315 by the 5th Amici prism 314, excited sample generation fluorescence 33 on the sample be positioned on displacement platform 318 is being radiated at through the 6th Amici prism 316 and microcobjective 317, fluorescent light beam carries out imaging by microcobjective 450, through the 6th Amici prism 316, optical filter 610, through fluorescence imaging lens combination 611, diaphragm 612 is imaged on EMCCD613.
So far, the microscope of the present embodiment transmission-type Quantization phase and fluorescence joint imaging is introduced complete.
It should be noted that, the above-mentioned definition to each element is not limited in the various concrete structure or shape mentioned in embodiment, and such as: rectangular parallelepiped glass lens can replace with wedge-shaped lens, microlens array can be terrace with edge also can be pyramid.
In sum, the present invention adopts dual wavelength light respectively through testee, the light with testee information formed interferes with the reference light through Spatial transmission, by CCD reception, record, and utilizes Spatial transmission structure to realize the separation of different wave length information.In the present invention, common focusing principle is applied in Quantization phase imaging, fluorescence imaging is combined with Quantization phase imaging, adopt the method for Spatial transmission and dual wavelength modulation, while the signal to noise ratio (S/N ratio) that improve Quantization phase microscopic system and resolution, obtain high-resolution fluoroscopic image, in cell fluorescence imaging, structure imaging, internal structure of body, Surface testing etc., there is good application, for cellular morphology and in site measurement provide a kind of new means.
Above-described specific embodiment; object of the present invention, technical scheme and beneficial effect are further described; be understood that; the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any amendment made, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (12)
1. the imaging of dual wavelength Quantization phase and a fluorescence imaging association system, is characterized in that, comprising:
Light-source system (100), for providing the first laser beam and second laser beam of different wave length;
Point tread assembly (200), for being the first reference beam and the first object light light beam by the first laser beam beam splitting; Be the second reference beam and the second object light light beam by the second laser beam beam splitting, the first reference beam and the second reference beam inject reference path (400); First object light light beam and the second object light light beam inject object light light path (300);
Object light light path (300), for by the first object light light beam and the second object light beam projection to sample, the first object light light beam and the second object light light beam that carry sample message reflex to interference imaging system (500) by the 5th Amici prism (314); Meanwhile, the fluorescent light beam produced by the first object light light beam and the second object light beam excitation sample is transmitted through fluoroscopic imaging systems (600) through the 6th Amici prism (316);
Reference path (400), for being projected to interference imaging system (500) by the first reference beam and the second reference beam;
Interference imaging system (500), for by the first reference beam and the first object light light beam, and the second reference beam and the second object light light beam carry out interference imaging;
Fluoroscopic imaging systems (600), shares co-focusing imaging part, for carrying out imaging to the fluorescent light beam of the first object light light beam and the second object light beam excitation with described object light light path (300).
2. dual wavelength Quantization phase according to claim 1 imaging and fluorescence imaging association system, it is characterized in that, described interference imaging system (500) comprising: imaging lens group (510), diaphragm (511) and interference imaging CCD (512), wherein:
First reference beam with carry the first object light light beam of sample message by imaging lens group (510) imaging, after diaphragm (511), interference imaging CCD (512) interferes, forms the interferogram with object information;
Second reference beam with carry the second object light light beam of sample message by imaging lens group (510) imaging, after diaphragm (511), interference imaging CCD (512) interferes, forms the interferogram with object information.
3. dual wavelength Quantization phase according to claim 1 imaging and fluorescence imaging association system, it is characterized in that, described reference path (400) comprising: the 4th half-wave plate (410), the second rectangular parallelepiped glass lens (411), the second Quantization phase adjuster (412) and the first catoptron (413); 3rd half-wave plate (420), the first rectangular parallelepiped glass lens (421), the first Quantization phase adjuster (422) and the 3rd Amici prism (423), wherein:
First reference beam is after the 3rd half-wave plate (420) and the first rectangular parallelepiped glass lens (421), adjust phase place by the first Quantization phase adjuster (422), and reflex to interference imaging system (500) through the 3rd Amici prism (423);
Second reference beam is after the 4th half-wave plate (410) and the second rectangular parallelepiped glass lens (411), phase place is adjusted by the second Quantization phase adjuster (412), through the first catoptron (413) reflection, and be transmitted through interference imaging system (500) by the 3rd Amici prism (423).
4. dual wavelength Quantization phase according to claim 3 imaging and fluorescence imaging association system, it is characterized in that, the phase modulator that described first Quantization phase adjuster (422) and the second Quantization phase adjuster (412) are double mirror type, it moves up and down the light path adjusting the first reference beam or the second reference beam by entirety.
5. dual wavelength Quantization phase according to claim 4 imaging and fluorescence imaging association system, it is characterized in that, described first rectangular parallelepiped glass lens (421) and the second rectangular parallelepiped glass lens (411) are rectangular parallelepiped glass lens, or substitute rectangular parallelepiped glass lens with wedge of glass lens.
6. dual wavelength Quantization phase according to claim 1 imaging and fluorescence imaging association system, it is characterized in that, described fluoroscopic imaging systems (600) comprising: microlens array (313), the 5th Amici prism (314), pinhole array (315), the 6th Amici prism (316), microcobjective (317) and displacement platform (318), optical filter (610), fluorescence imaging lens combination (611), diaphragm (612) and EMCCD (613), wherein:
First object light light beam and the second object light light beam irradiation are in microlens array (313), produce the array with certain space periodic arrangement, this array is mated with pinhole array (315) by the 5th Amici prism (314), excited sample generation fluorescence on the sample be positioned on displacement platform (318) is being radiated at through the 6th Amici prism (316) and microcobjective (317), fluorescent light beam carries out imaging by microcobjective (317) again through displacement platform (318) reflection, through the 6th Amici prism (316), optical filter (610), through fluorescence imaging lens combination (611), diaphragm (612) is imaged on EMCCD (613).
7. dual wavelength Quantization phase according to claim 6 imaging and fluorescence imaging association system, is characterized in that, substitutes described microlens array with terrace with edge or pyramid.
8. dual wavelength Quantization phase according to any one of claim 1 to 7 imaging and fluorescence imaging association system, it is characterized in that, described light-source system (100) comprising: the first laser instrument (110), the first beam expander (111), the first half-wave plate (112), second laser (120), the second beam expander (121), the second half-wave plate (122), wherein:
First laser instrument (110) sends the first laser beam of first wave length, this first laser beam carries out beam-expanding collimation through the first beam expander (111), then enters beam splitting assembly (200) after the first half-wave plate (112);
Second laser (120) sends the second laser beam of second wave length, this second laser beam carries out beam-expanding collimation through the second beam expander (121), then enters beam splitting assembly (200) after the second half-wave plate (122).
9. dual wavelength Quantization phase according to claim 8 imaging and fluorescence imaging association system, it is characterized in that, described first wave length is the one in 346nm, 495nm, 514nm, 556nm, 647nm or 710nm, and second wave length is the another kind in 346nm, 495nm, 514nm, 556nm, 647nm or 710nm.
10. dual wavelength Quantization phase according to any one of claim 1 to 7 imaging and fluorescence imaging association system, it is characterized in that, described point of tread assembly (200) comprises the first polarization splitting prism (211) and the second polarization splitting prism (221) that mutually stagger, wherein:
First laser beam by the first polarization splitting prism (211) light splitting be along the horizontal plane direction propagate the first object light light beam and along vertical plane direction propagate the first reference light light beam;
Second laser beam by the second polarization splitting prism (221) light splitting be along the horizontal plane direction propagate the second object light light beam and along vertical plane direction propagate the second reference light light beam.
11. dual wavelength Quantization phase according to any one of claim 1 to 7 imagings and fluorescence imaging association system, it is characterized in that, described object light light path (300) comprising: the second catoptron (311), the 4th Amici prism (312), microlens array (313), the 5th Amici prism (314), pinhole array (315), the 6th Amici prism (316), microcobjective (317) and displacement platform (318), wherein:
First object light light beam is after the second catoptron (311) reflection, through the 4th Amici prism (312) transmission, be radiated at microlens array (313), thus produce the array with certain space periodic arrangement, this array is mated with pinhole array (315) by the 5th Amici prism (314), be radiated on the sample be positioned on displacement platform (318) through the 6th Amici prism (316) and microcobjective (317), thus become the first object light light beam carrying sample message, imaging is carried out by microcobjective (317) again through displacement platform (318) reflection, through the 6th Amici prism (316), elimination veiling glare is being mated with pinhole array (315), finally be projected to the 5th Amici prism (314), through the 5th Amici prism (314) catoptric imaging to interference imaging system (500),
Second object light light beam is after the 4th Amici prism (312) reflection, be radiated at microlens array (313), thus produce the array with certain space periodic arrangement, this array is mated with pinhole array (315) by the 5th Amici prism (314), be radiated on the sample be positioned on displacement platform (318) through the 6th Amici prism (316) and microcobjective (317), thus become the first object light light beam carrying sample message, imaging is carried out by microcobjective (317) again through displacement platform (318) reflection, through the 6th Amici prism (316), elimination veiling glare is being mated with pinhole array (315), finally be projected to the 5th Amici prism (314), through the 5th Amici prism (314) catoptric imaging to interference imaging system (500).
12. dual wavelength Quantization phase according to claim 11 imagings and fluorescence imaging association system, is characterized in that, substitutes described microlens array with terrace with edge or pyramid.
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