CN106124472A - A kind of face battle array detecting stimulated radiation loss imaging system - Google Patents
A kind of face battle array detecting stimulated radiation loss imaging system Download PDFInfo
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- CN106124472A CN106124472A CN201610594017.3A CN201610594017A CN106124472A CN 106124472 A CN106124472 A CN 106124472A CN 201610594017 A CN201610594017 A CN 201610594017A CN 106124472 A CN106124472 A CN 106124472A
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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
- G01N21/62—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
- G01N21/645—Specially adapted constructive features of fluorimeters
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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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
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Abstract
nullA kind of face battle array detecting stimulated radiation loss imaging system，Including the exciting light sources for producing exciting light、For producing the loss radiant of loss light、Beam-expanding system、Vortex phase modulation panel、Dichroscope、Wave plate、Scanning galvanometer、Scanning lens、Cylinder mirror、Object lens、Nanometer displacement platform、Focal length collective lens and planar array detector，Exciting light after described beam-expanding system expands with through described beam-expanding system、Vortex phase modulation panel carry out expanding and loss light after phase-modulation enter the reflection of described dichroscope after enter described wave plate，By the exciting light of described wave plate with loss light after described scanning galvanometer，Pass sequentially through through described scanning lens、After cylinder mirror and object lens，Focus on the sample interior at described nanometer displacement platform，Focus on the exciting light of the sample interior at described nanometer displacement platform and the optical signal of loss light generation by detecting light path、Focal length collective lens converges at imaging on described planar array detector.The application is easy and simple to handle, the efficiency of light energy utilization is high.
Description
Technical field
The application relates to super-resolution fluorescence micro-imaging field.
Background technology
Tradition far-field optics microscopic resolution is mainly limited by diffraction limit, its focus formed at object focal point
Size, i.e. point spread function (PSF) determine its resolution capability.Stimulated radiation loss (Stimulated emission
Depletion, STED) imaging technique is a kind of far-field optics super-resolution imaging technology, it breaches traditional optical diffraction limit
Restriction, far-field optics resolution improve the highest to 50nm.
Stimulated radiation loss image-forming principle is the laser using two-way wavelength different, and a road laser is called exciting light, is used for
In exciting the samples such as biology, fluorescent material is to excited state, thus gives off detectable fluorescence；Another road laser is called loss light,
Same purpose, in the fluorescent material being excited, can make it return ground state in excited state, but this process not radiofluorescence, but
Radiate the light identical with loss light, it is possible to filter from detection light path.By the use of light is lost, can control glimmering artificially
Light radiation, reaches the effect of photoswitch, it is achieved the reversible switching of fluorescence open/close state.
In imaging system, loss light path have employed 0-2 π vortex phase modulation loss light wavefront so that it is put down object lens Jiao
Forming the annular focus (i.e. bagel hot spot) of a hollow at face, the solid circles hot spot that this hot spot and exciting light are formed is empty
Between overlap.So, the hollow position of hot spot central authorities is not lost the impact of light, fluorescence molecule can with normal radiation fluorescence,
And the fluorescence molecule of periphery is acted on by loss light, create the state that fluorescence is closed so that effective excitation area is pressed down
System is within annulus hot spot.So, it is possible to directly reduce the size of object focal point PSF, thus improve imaging resolution.Class
Like co-focusing imaging, use the mode imaging of point by point scanning, thus obtain the fluorescence intensity profile picture that sample is two-dimentional or three-dimensional.
But, in detection side, mostly use point probe device, use pinhole filter before the detectors, or use
The end face of multimode fibre substitutes pin hole, and aperture size is traditionally arranged to be about 1 times of Airy disk diameter, below the detection existence of pin hole point
Problem:
Debuging difficulty: two-way laser needs at space coincidence, the maximum of exciting light needs with loss light hollow minima
Strict coincidence, could realize high-resolution imaging.If coincidence deviation is relatively big, then imaging resolution and signal to noise ratio all significantly reduce.
Point detection system cannot monitor the degree that two-way laser overlaps in real time, typically need to use gold particle imaging, separately detect two-way
The mode of laser PSF carries out the conjunction bundle of precision, before actual sample imaging, needs to be replaced by gold particle sample, and to goldc grains increment
The repetitious scanning imagery of product determine two-way laser relative to position, and fine adjustment, operating process is extremely complex.Additionally, be
In system, pin hole position is if the deviation from primary optical axis, will also result in the drastically decline of signal intensity, it is therefore desirable to keep pin hole and key light
The height of axle overlaps, but owing to pinhole diameter only has 1 times of Airy disk size (50-100 micron), it is easy to former due to machinery etc.
Deviateed by generation, cause system imaging Quality Down, need before each imaging to adjust pin hole position, add the complexity of operation
Property.
Energy loss: stimulated radiation loss belongs to low light level imaging, and signal value is the lowest, and the pin hole meeting of 1 times of Airy disk size
Stop passing through of fluorescence in space, cause the further decline of the efficiency of light energy utilization, be unfavorable for weak light detection, had a strong impact on one-tenth
Picture element amount.
Sensing point spread function is fixed: employ pinhole detector in system, then the sensing point spread function of this system is solid
Determine to get off, there is no the leeway that can regulate, it is impossible to start with in terms of sensing point spread function, improved the one-tenth of stimulated radiation loss
Picture element amount.
Summary of the invention
In consideration of it, be necessary that offer one is easy and simple to handle, the efficiency of light energy utilization is high, adjustable battle array of sensing point spread function is visited
Survey type stimulated radiation loss imaging system.
For solving above-mentioned technical problem, this application provides a kind of face battle array detecting stimulated radiation loss imaging system, bag
Include the exciting light sources for producing exciting light, for producing the loss loss radiant of light, beam-expanding system, vortex phase modulation
Plate, dichroscope, wave plate, scanning galvanometer, scanning lens, cylinder mirror, object lens, nanometer displacement platform, focal length collective lens and the detection of face battle array
Device, the exciting light after described beam-expanding system expands carries out expanding and phase place with through described beam-expanding system, vortex phase modulation panel
Loss light after modulation enters described wave plate after entering the reflection of described dichroscope, by the exciting light of described wave plate and loss light warp
After crossing described scanning galvanometer, pass sequentially through after described scanning lens, cylinder mirror and object lens, focus at described nanometer displacement platform
Sample interior, focus on the optical signal that the exciting light of the sample interior at described nanometer displacement platform and loss light produces and pass through institute
State focal length collective lens and converge at imaging on described planar array detector.
Preferably, described beam-expanding system includes the first beam-expanding system, the second beam-expanding system and the 3rd beam-expanding system, described
Dichroscope includes the first dichroscope and the second dichroscope.
Preferably, the exciting light that described exciting light sources produces expands through described first beam-expanding system.
Preferably, described loss radiant produce loss light after first, second beam-expanding system expands again through described
Vortex phase modulation panel carries out phase-modulation.
Preferably, the exciting light through expanding passes through described first, second 2 colors with via the loss light after phase-modulation
Enter in described wave plate after mirror reflection.
Preferably, described scanning galvanometer be used for scanning or described nanometer displacement platform for scan or described scanning galvanometer with
The cooperation of described nanometer displacement platform is used for scanning.
Preferably, described wave plate is quarter wave plate.
The application face battle array detecting stimulated radiation loss imaging system, compared to tradition stimulated radiation loss imaging system,
Actually achieving the regulation to sensing point spread function, described planar array detector can be according to practical situation needs, opposite
Each picture dot of array detector reads data independently, such that it is able to independent, freely regulate search coverage size,
The parameter such as position, gain, such that it is able to improve image quality.For example, it is possible to realize the effect of dummy pinhole, utilize described battle array
The actual pin hole effect of the search coverage virtual tradition stimulated radiation loss imaging system of the picture dot composition of detector, and according to choosing
The picture dot area size selecting data streams read is different, can be advantageously converted into the size of dummy pinhole.
Need to use complicated gold particle to carry out an expansion compared to existing some detecting stimulated radiation loss imaging system
Dissipating the measurement of function, battle array detecting stimulated radiation loss imaging system in the application face uses planar array detector to detect, permissible
The reflection light position on planar array detector of convenience, real-time monitoring and regulation exciting light and loss light, thus realize high-precision
Degree closes the bundle visualization of regulation process, easy.And can effectively reduce the deviation effects that detection light path is caused the most off axis,
The application substantially increases the stability of total system and the simplicity of operation.Described planar array detector have collected all fluorescence energy
Amount, improves the efficiency of light energy utilization, and this, for belonging to weak light detection stimulated radiation loss imaging system, has the most great
Meaning.
Accompanying drawing explanation
Accompanying drawing described herein is used for providing further understanding of the present application, constitutes the part of the application, this Shen
Schematic description and description please is used for explaining the application, is not intended that the improper restriction to the application.In the accompanying drawings:
Fig. 1 is the schematic diagram of the application face battle array detecting stimulated radiation loss imaging system；
Fig. 2 is the face battle array detection schematic diagram of the application face battle array detecting stimulated radiation loss imaging system；
Fig. 3 is that signal when restrainting deviation is closed in the face battle array detection of the application face battle array detecting stimulated radiation loss imaging system
Figure；
Fig. 4 be the planar array detector center of the application face battle array detecting stimulated radiation loss imaging system off-axis time signal
Figure.
Detailed description of the invention
For making the purpose of the application, technical scheme and advantage clearer, below in conjunction with the application specific embodiment and
Technical scheme is clearly and completely described by corresponding accompanying drawing.Obviously, described embodiment is only the application one
Section Example rather than whole embodiments.Based on the embodiment in the application, those of ordinary skill in the art are not doing
Go out the every other embodiment obtained under creative work premise, broadly fall into the scope of the application protection.
Below in conjunction with accompanying drawing, describe the technical scheme that each embodiment of the application provides in detail.
Referring to shown in Fig. 1, battle array detecting stimulated radiation loss imaging system in the application face includes excitation source 1, loss
Radiant 2, beam-expanding system 3, vortex phase modulation panel 5, dichroscope 4, wave plate 6, scanning galvanometer 7, scanning lens 8, cylinder mirror 9, thing
Mirror 10, nanometer displacement platform 11, focal length collective lens 12 and planar array detector 13.Described beam-expanding system 3 includes the first beam-expanding system
3-1, the second beam-expanding system 3-2 and the 3rd beam-expanding system 3-3.Described dichroscope 4 includes the first dichroscope 4-1 and the two or two color
Mirror 4-2.Described nanometer displacement platform 11 is used for carrying scanning sample.
Described exciting light sources 1 produces exciting light and expands through described first beam-expanding system 3-1.Described loss light
Light source 2 produces loss light and enters through described vortex phase modulation panel 5 after first, second beam-expanding system 3-2,3-3 expands again
Line phase is modulated.
The exciting light expanded via described first beam-expanding system 3-1 with via described vortex phase modulation panel 5 phase-modulation
After loss light all enter described first, second dichroscope 4-1,4-2 reflection after enter in described wave plate 6.By described ripple
The exciting light of sheet 6 and loss light, after described scanning galvanometer 7, pass sequentially through through described scanning lens 8, cylinder mirror 9 and object lens
After 10, focus on the sample interior at described nanometer displacement platform 11.The scan mode of the application can use described scanning galvanometer 7
It is scanned, it is possible to use described nanometer displacement platform 11 is scanned.
The optical signal of the exciting light and loss light generation that focus on the sample interior at described nanometer displacement platform 11 then passes through
Described focal length collective lens 12 converges on described planar array detector 13.The detection plane of described planar array detector 13 is displaced into institute
State on the focal plane of focal length collective lens 12.
Described wave plate 6 is quarter wave plate.Described planar array detector 13 is collected fluorescence by described focal length collective lens 12 and is believed
Number.
As in figure 2 it is shown, wherein, label 14 represents that exciting light is in face to the test surface imaging schematic diagram of described planar array detector 13
The hot spot formed on detector, label 15 represents the hot spot that loss light is formed on surface detector.Grid shown in label 16 represents
Some picture dots on planar array detector.
During regulation light path, Imaging for Monitoring situation that can be real-time by described surface detector 13, determine exciting light
With loss light center position on described planar array detector 13, thus realize the high-precision of two-way laser (exciting light and loss light)
Degree, easy conjunction are restrainted, and make regulation process intuitively visualize.And when exciting light exists deviation with loss combiner, described battle array is visited
Survey the imaging effect on the CCD of device 13 and make exciting light consistent with loss combiner as it is shown on figure 3, now need to regulate light path,
Visualize according to the CCD screen of described planar array detector 13 during regulation light path and carry out, easy and simple to handle.
The detection area of described planar array detector 13 is much larger than one times of Airy disk, and all luminous energy are detected by described battle array completely
Device 13 receives, it is not necessary to worry the impact that detection light path is caused the most off axis, it is only necessary to according to practical situation, picture dot is read in regulation
Position, the described planar array detector 13 effect when detecting light path and being slightly off-axis is as shown in Figure 4, although exciting light is in face
The hot spot 15 that the hot spot 14 formed on array detector 13 and loss light are formed on planar array detector 13 not in precalculated position, but its
Still completely can display on screen, without causing optical energy loss.
The application face battle array detecting stimulated radiation loss imaging system, compared to tradition stimulated radiation loss imaging system,
Actually achieving the regulation to sensing point spread function, described planar array detector 13 can be right according to practical situation needs
Each picture dot of planar array detector 13 reads data independently, such that it is able to independently, freely regulate search coverage
The parameters such as size, position, gain, such that it is able to improve image quality.For example, it is possible to realize the effect of dummy pinhole, utilize institute
State the actual pin hole effect of the search coverage virtual tradition stimulated radiation loss imaging system of the picture dot composition of planar array detector 13,
And different according to the picture dot area size selecting data streams read, the size of dummy pinhole can be advantageously converted into.
Need to use complicated gold particle to carry out an expansion compared to existing some detecting stimulated radiation loss imaging system
Dissipating the measurement of function, battle array detecting stimulated radiation loss imaging system in the application face uses planar array detector 13 to detect, can
With the reflection light position on planar array detector of convenient, real-time monitoring and regulation exciting light and loss light, thus realize height
Precision closes the bundle visualization of regulation process, easy.And can effectively reduce the deviation shadow that detection light path is caused the most off axis
Ringing, the application substantially increases the stability of total system and the simplicity of operation.Described planar array detector 13 have collected all
Fluorescent energy, improves the efficiency of light energy utilization, and this, for belonging to weak light detection stimulated radiation loss imaging system, has very
Great meaning.
Those skilled in the art are it should be appreciated that embodiments of the invention can be provided as method, system or computer program
Product.Therefore, the reality in terms of the present invention can use complete hardware embodiment, complete software implementation or combine software and hardware
Execute the form of example.And, the present invention can use at one or more computers wherein including computer usable program code
The upper computer program product implemented of usable storage medium (including but not limited to disk memory, CD-ROM, optical memory etc.)
The form of product.
Also, it should be noted term " includes ", " comprising " or its any other variant are intended to nonexcludability
Comprise, so that include that the process of a series of key element, method, commodity or equipment not only include those key elements, but also wrap
Include other key elements being not expressly set out, or also include want intrinsic for this process, method, commodity or equipment
Element.In the case of there is no more restriction, statement " including ... " key element limited, it is not excluded that including described wanting
Process, method, commodity or the equipment of element there is also other identical element.
The foregoing is only embodiments herein, be not limited to the application.For those skilled in the art
For, the application can have various modifications and variations.All made within spirit herein and principle any amendment, equivalent
Replacement, improvement etc., within the scope of should be included in claims hereof.
Claims (7)
1. a face battle array detecting stimulated radiation loss imaging system, it is characterised in that include for producing exciting of exciting light
Radiant, for producing the loss loss radiant of light, beam-expanding system, vortex phase modulation panel, dichroscope, wave plate, scanning are shaken
Mirror, scanning lens, cylinder mirror, object lens, nanometer displacement platform, focal length collective lens and planar array detector, expand through described beam-expanding system
After exciting light carry out expanding and loss light after phase-modulation enters described with through described beam-expanding system, vortex phase modulation panel
Dichroscope reflection after enter described wave plate, by the exciting light of described wave plate with loss light after described scanning galvanometer, successively
By after described scanning lens, cylinder mirror and object lens, focus on the sample interior at described nanometer displacement platform, focus on described
The optical signal that the exciting light of the sample interior at nanometer displacement platform produces with loss light is converged at by described focal length collective lens
Imaging on described planar array detector.
2. face as claimed in claim 1 battle array detecting stimulated radiation loss imaging system, it is characterised in that described beam-expanding system
Including the first beam-expanding system, the second beam-expanding system and the 3rd beam-expanding system, described dichroscope includes the first dichroscope and the two or two
Color mirror.
3. face as claimed in claim 2 battle array detecting stimulated radiation loss imaging system, it is characterised in that described excitation light
The exciting light that source produces expands through described first beam-expanding system.
4. face as claimed in claim 2 battle array detecting stimulated radiation loss imaging system, it is characterised in that described loss light light
The loss light that source produces carries out phase-modulation through described vortex phase modulation panel after first, second beam-expanding system expands again.
5. face as claimed in claim 2 battle array detecting stimulated radiation loss imaging system, it is characterised in that through swashing of expanding
Luminescence is entered in described wave plate after being reflected by described first, second dichroscope via the loss light after phase-modulation.
6. face as claimed in claim 1 battle array detecting stimulated radiation loss imaging system, it is characterised in that described scanning galvanometer
For scanning or described nanometer displacement platform is for scanning or described scanning galvanometer coordinates for scanning with described nanometer displacement platform.
7. face as claimed in claim 1 battle array detecting stimulated radiation loss imaging system, it is characterised in that described wave plate is 1/
4 wave plates.
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Cited By (6)
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| CN107941763A (en) * | 2017-10-27 | 2018-04-20 | 浙江大学 | A kind of coaxial three-dimensional stimulated radiation loss super-resolution micro imaging method and device |
| CN109062268A (en) * | 2018-08-21 | 2018-12-21 | 中国科学院电工研究所 | A kind of nanometer displacement platform scan time delay system and its implementation |
| CN110596059A (en) * | 2019-09-05 | 2019-12-20 | 北京世纪桑尼科技有限公司 | Optical super-resolution microscopic imaging system |
| CN113566745A (en) * | 2021-07-30 | 2021-10-29 | 上海无线电设备研究所 | High-precision roll angle measuring device and method based on laser collimation technology |
| WO2022253098A1 (en) * | 2021-05-31 | 2022-12-08 | 苏州德龙激光股份有限公司 | Laser scanning microscopic measurement device and method |
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| CN113566745B (en) * | 2021-07-30 | 2024-02-20 | 上海无线电设备研究所 | High-precision roll angle measuring device and method based on laser collimation technology |
| CN116300310A (en) * | 2023-01-06 | 2023-06-23 | 之江实验室 | Method and device for realizing super-resolution inscription and imaging by utilizing photoinitiator |
| CN116300310B (en) * | 2023-01-06 | 2024-04-16 | 之江实验室 | Method and device for realizing super-resolution inscription and imaging by utilizing photoinitiator |
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Application publication date: 20161116 |