CN208140563U - Microscopic system is just set in up-conversion luminescence near-infrared and the multi-functional wide field of colour imaging - Google Patents
Microscopic system is just set in up-conversion luminescence near-infrared and the multi-functional wide field of colour imaging Download PDFInfo
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- CN208140563U CN208140563U CN201820646690.1U CN201820646690U CN208140563U CN 208140563 U CN208140563 U CN 208140563U CN 201820646690 U CN201820646690 U CN 201820646690U CN 208140563 U CN208140563 U CN 208140563U
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Abstract
The utility model discloses a kind of up-conversion luminescence near-infrareds and the multi-functional wide field of colour imaging just to set microscopic system, the microscopic system successively integrally disposed from the bottom to top in fluorescence microscope mirror body has the penetrating object lens of near-infrared just setting, exciter filter turntable, dichroscope, emit filter wheel, second adjustable reflecting mirror and high sensitivity detector, color camera is provided with just setting the side in fluorescence microscope mirror body positioned at the second adjustable reflecting mirror, the first adjustable reflecting mirror is sequentially installed in the side that the outside for just setting fluorescence microscope mirror body is located at installation dichroscope position, collimator and extender mirror and laser, white light source is set to the side of the first adjustable reflecting mirror.This system is excited by near infrared light, emitted using the visible light or near infrared light of highly-sensitive detector detection probe, the colour imaging of sample, the final material for realizing probe, cell or real-time, non-intrusion type the highly sensitive imaging of biological tissue can be realized by switching simultaneously.
Description
Technical field
The utility model belongs to Imaging-PAM field, particularly relates to a kind of up-conversion luminescence near-infrared and coloured silk
Color is imaged multi-functional wide field and just sets microscopic system.
Background technique
Optical microscopy is a kind of microscope that image zoom effect is generated using optical lens, and just setting microscope is optics
Microscopical one kind, under viewed in transmitted light, light source reaches sample by passing through condenser below fuselage, is then passed through on sample
Then the object lens of side reach the eyes or other Image-forming instruments of observer by reflecting mirror and lens, are just setting microscopical advantage
It is that structure is simple, is easy to be transformed.Induced Fluorescence Microscopy refers to using fluorescence the technology for generating an image.Sample is shone
The light for penetrating specific wavelength or wave band, is absorbed by fluorogen, cause they issue more long wavelengths light, such as with absorbed light
Different colors.By using spectral emissions filter disc, which is separated from much weaker transmitting fluorescence.In recent years
Come, in biological study, fluorescence labels are widely used to demarcate biomolecule, and fluorescence microscope is made to become more to aggravate
It wants, fluorescence microscope is collocation tool excitation filter disc, the optical instrument for dissipating filter disc group using mercury vapor lamp or xenon lamp as light source.It is glimmering
Light micro imaging system is widely used in the research of cell biology, is indispensable one of the tool of cellular level research.
Near-infrared fluorescence imaging belongs to one kind of optical imagery, and the wave-length coverage of near-infrared light source is 700~900 nm, is
The non-visible region that people have found earliest, in the imaging of 400~700 nm of visible region, there is many problems, such as can be by
The endogenous material into biological tissue, such as absorption, the scattering etc. pair of aerobic, anaerobic hemoglobin, melanin, water and bilirubin
The influence of optical imagery.Then in the near infrared region, the absorption, scattering of tissue and autofluorescence background are all very low, near infrared light
Source can reach maximum penetration in biological tissue, and can be carried out deep tissues imaging, thus this wavelength band is referred to as " close red
Outer transparency of organization window ".Up-conversion fluorescence imaging is based on up-conversion fluorescence probe, other fluorescence probes is different from, in long wave
The lower fluorescence that can emit short wavelength of long external light source excitation, this unique luminosity make up-conversion fluorescence probe exist
When carrying out fluorescence imaging, the autofluorescence of biological tissue can be avoided to interfere significantly, and the light of up-conversion fluorescence probe is stablized
Property it is also very high, it is not easy to the problems such as fluorescent quenching occurs, suitable for organism long-term fluorescence imaging observe.
Fluorescence microscopy system its cost relative moderate, image taking speed be fast, high sensitivity, and near infrared light and up-conversion luminescence are equal
Have the advantages that imaging depth is good, low noise, be based on above-mentioned advantage, development can be realized the aobvious of near infrared light and up-conversion luminescence
Micro- imaging microscopic system is very necessary, simultaneously because the needs with the pathological examinations common location such as HE dyeing, simple switching are realized
The colour imaging of original position, near-infrared luminous and up-conversion luminescence imaging have a very important significance the research of cellular level.
Summary of the invention
The utility model in order to overcome the shortcomings of the prior art, provide a kind of up-conversion luminescence near-infrared and it is colored at
As microscopic system is just set in multi-functional wide field.
The utility model is achieved through the following technical solutions：A kind of up-conversion luminescence near-infrared and the more function of colour imaging
Microscopic system is just set in energy wide field, which is just setting microscopic system mainly by just setting fluorescence microscope mirror body, laser, standard
Straight beam expanding lens, white light source, the first adjustable reflecting mirror, dichroscope, exciter filter turntable, the penetrating object lens of near-infrared, hair
Penetrate filter wheel, the second adjustable reflecting mirror, high sensitivity detector and color camera composition；Just setting fluorescence microscope mirror
It is successively integrally disposed from the bottom to top in vivo to there is the penetrating object lens of near-infrared, exciter filter turntable, dichroscope, transmitting optical filter to turn
Disk, the second adjustable reflecting mirror and high sensitivity detector are located at the second adjustable reflection just setting in fluorescence microscope mirror body
The side of mirror is provided with color camera, the outside for just setting fluorescence microscope mirror body be located at installation dichroscope position side according to
Secondary to be equipped with the first adjustable reflecting mirror, collimator and extender mirror and laser, white light source is set to the first adjustable reflecting mirror
Top.
Exciter filter turntable contains exciter filter, and transmitting filter wheel contains transmitting light optical filter, and excitation filters
Piece and transmitting light optical filter use short pass filter, bandpass filter or long pass filter.
Laser uses central wavelength for the laser of 980nm, 808nm or 730nm.White light source uses mercury lamp or LED
Lamp.High sensitivity detector is using sCMOS detector or refrigeration property EMCCD detector.
The function that the utility model is just setting each component in microscopic system is：
1, just setting that fluorescence microscope mirror body is connected as each component and physics is fixed.
2, excitation light source of the laser as near-infrared luminous imaging and up-conversion luminescence imaging.
3, collimator and extender mirror is used for the collimation of laser and expands.
4, transmitting light source of the white light source as exciting light.
5, the first adjustable reflecting mirror can switch over light source, switched laser light source and white light source.
6, dichroscope can excite it to sample with reflected excitation light, while can make to emit through transmitting light
Light can be received by detector.
7, exciter filter turntable contains exciter filter, exciter filter be capable of selectivity through laser, mercury lamp,
LED light source emits light as exciting light.
8, the penetrating object lens of near-infrared are exclusively used in the lens group of infrared light.
9, transmitting filter wheel contain transmitting light optical filter, transmitting light optical filter be capable of selectivity penetrate objective emission
Light.
10, the second adjustable reflecting mirror can be used to implement freely cutting between high sensitivity detector and color camera
It changes.
11, high sensitivity detector is used to collect the fluorescence emitted after sample excitation.
12, color camera is used for the colour imaging of visible light wave range.
Just set fluorescence microscope：Nikon Ni-U is that the extremely strong scientific research grade of a scalability just sets microscope, using unlimited
Remote optical system, it is convenient to carry other attachmentes.It is equipped with the double-deck fluorescence turntable in such a system, every layer of fluorescence turntable has
Six optical filtering block vacancy, system are further provided with double camera port and double light source interfaces, with simultaneously meet biological fluorescence material and on
The application of changing luminous material.On major optical component, as object lens select near-infrared transmitance is higher to be greater than 70%
Flat field fluorescence object lens.In addition under the premise of ensuring does not influence imaging, the optics for having barrier effect in optical path to infrared light is removed
Component, to improve entire optical system to the maximum extent in the percent of pass of near infrared light.
Laser uses the big core diameter multimode fibre of high power, to reach stronger excitating light strength.Optical fiber uses multimode light
Fibre, core diameter 200um, additional collimator and extender mirror realize uniform field illumination in turn.Camera in addition to the combination of accurately optical filter and
Outside the object lens of high transmittance, the fluorescent image of high s/n ratio, system also need to be equipped with highly sensitive detector in order to obtain, such as
EMCCD or sCMOS.
Dichroscope is different from the dichroscope of common fluorescent, and using in the utility model is short flow-through optical filter,
That is the light of shorter than light splitting wavelength penetrates, and the light for being longer than light splitting wavelength is reflected.After the near infrared light to sample reflected,
The fluorescence that sample excitation goes out allows the light of 400nm~850nm to penetrate, the light of commplementary wave length is filtered by the first barrier filter
Fall, avoids interfering background signal.It is required that dichroscope and first stop optical filter OD value at least more than 6, with increase
Signal-to-noise ratio.Microscopic system is just set using the ZT1064rdc-sp dichroscope of Chroma company of the U.S. in the utility model
And ET850SP-2P, for spectrogram as shown in Fig. 2, wherein ZT1064rdc-sp is the smooth shape line of grey, ET850SP-2P is black
Jagged line.
Excitation and transmitting optical filter：In order to further discriminate between out the transmitting light of different wave length, the second resistance is increased in system
Optical filter is kept off, wavelength is respectively ET550/50M, ET630/75M and ET811/80M, spectrogram such as Fig. 3, Fig. 4 and Fig. 5, wherein
Black line is the first barrier filter.Further, since conventional fluorescence microscope light source is using mercury lamp, heat is relatively high,
So optical path usually has heat shield.And heat shield can be by near-infrared and infrared light blocking, so this system is to fluorescent
Related optical component in optical path is also removed.Exciter filter turntable and transmitting filter wheel can arbitrarily replace filter
Mating plate can also realize the free switching of optical filter in imaging process.
The utility model has the beneficial effects that：The utility model provides a kind of up-conversion luminescence near infrared imaging and colour
The characteristics of micro imaging system, which can be using near infrared light, up-conversion luminescence signal-to-noise ratio height, low noise, by close red
Outer light excitation, is emitted using the visible light or near infrared light of highly-sensitive detector detection probe, while can be real by switching
The colour imaging of existing sample, final material, cell or real-time, non-intrusion type the high sensitivity of biological tissue for realizing probe
Imaging.Microscopic system in the utility model can emit exciting light, by having probe, near-infrared fluorescent to label
Probe, the material of up-conversion luminescence probe, cell, tissue and living animal carry out the fluorescence microscopy of real-time high s/n ratio at
Picture, while common location colour imaging can be carried out using color camera in the case where not moving sample and moving lens.This reality
With novel by reasonably combined filter spectral range, the micro objective and high OD value optical filter of high transmittance are selected, simultaneously
It is equipped with highly sensitive detector, equally can achieve the up-conversion fluorescence image of high s/n ratio under Both wide field illumination.
The utility model is based on just setting microscope, by the optical component of modernization system, increases visible light and near infrared light
Highly sensitive detector and color imaging system increase superpower laser, and increase and match high-efficient filter mating plate system, construct a set of energy
Enough carry out upper conversion imaging, near infrared imaging and colour imaging just sets microscopic system.The utility model and it is previous it is micro- at
As system is compared, there is highly sensitive and high s/n ratio, be one kind be able to carry out visible light, near infrared light and up-conversion luminescence at
The integrated micro-imaging equipment of picture.
Compared with prior art, the imaging system in the utility model has further the advantage that：1, realizing for the first time will be general
Logical fluorescence imaging, near-infrared micro-imaging technique, upper conversion living body fluorescent micro-imaging technique and colour image technique all collect
At into a microscopic system；2, can in imaging process free switching to common fluorescent imaging pattern, near infrared light imaging
Mode, up-conversion luminescence imaging pattern and colour imaging mode, to realize the common location of different imaging results；3, near-infrared is aobvious
Micro- imaging and upper conversion living body fluorescent micro-imaging have higher signal-to-noise ratio.
Detailed description of the invention
Fig. 1 is the system light path schematic diagram that the utility model is just setting microscopic system；
Fig. 2 is the spectrogram of dichroscope and the first barrier filter in the utility model；
Fig. 3 is the ET550/50m filter spectral figure of the utility model；
Fig. 4 is the ET630/75m filter spectral figure of the utility model；
Fig. 5 is the ET811/80m filter spectral figure of the utility model；
Fig. 6 is just to set the high s/n ratio up-conversion fluorescence image that microscopic system obtains using the utility model；
In figure：1- is just setting fluorescence microscope mirror body；2- laser；3- collimator and extender mirror；4- white light source；5- first is adjustable
Formula reflecting mirror；6- dichroscope；7- exciter filter turntable；The penetrating object lens of 8- near-infrared；9- emits filter wheel；10-
Two adjustable reflecting mirrors；11- high sensitivity detector；12- color camera.
Specific embodiment
The utility model is described in detail below in conjunction with the drawings and specific embodiments.
As shown in Figures 1 to 6, microscopic system is just set in a kind of up-conversion luminescence near-infrared and the multi-functional wide field of colour imaging,
Microscopic system is just being set mainly by just setting fluorescence microscope mirror body 1, laser 2, collimator and extender mirror 3, white light light in the multi-functional wide field
Source 4, the first adjustable reflecting mirror 5, dichroscope 6, exciter filter turntable 7, the penetrating object lens 8 of near-infrared, transmitting optical filter turn
Disk 9, the second adjustable reflecting mirror 10, high sensitivity detector 11 and color camera 12 form；Just setting fluorescence microscope mirror body 1
It is inside successively integrally disposed from the bottom to top to have the penetrating object lens 8 of near-infrared, exciter filter turntable 7, dichroscope 6, transmitting optical filter
Turntable 9, the second adjustable reflecting mirror 10 and high sensitivity detector 11, can positioned at second in fluorescence microscope mirror body 1 just setting
The side of mode reflecting mirror 10 is provided with color camera 12, is located at installation dichroic in the outside for just setting fluorescence microscope mirror body 1
The side of 6 position of mirror is sequentially installed with the first adjustable reflecting mirror, collimator and extender mirror 3 and laser 2, and white light source 4 is set to
The side of first adjustable reflecting mirror 5.
Exciter filter turntable 7 contains exciter filter, and transmitting filter wheel 9 contains transmitting light optical filter, excitation filter
Mating plate and transmitting light optical filter use short pass filter, bandpass filter or long pass filter.Laser 2 use central wavelength for
The laser 2 of 980nm, 808nm or 730nm.White light source 4 uses mercury lamp or LED light.High sensitivity detector 11 uses
SCMOS detector or refrigeration property EMCCD detector.
It includes just setting fluorescence microscope mirror body 1, colour+forming monochrome image dual-port, white light that the system of the utility model, which is constituted,
2 pairs of light source interfaces of LED+ laser, the double-deck fluorescence optical filtering block turntable and a series of excitations for realizing function and transmitting optical filter
Composition.This system is based on one and is just setting fluorescence microscope Nikon Ni-U, and light source is imported using the optical fiber of 980nm continuous laser,
By equably injecting microscopes optical path after collimator and extender, after the light splitting of dichroscope 6, it is reflected through object lens and is irradiated to sample
On product, after the fluorescence and unabsorbed exciting light that sample generates return to the light splitting of dichroscope 6 by object lens, high sensitivity is used
Camera receives optical signal, is handled after digitlization by computer software.Color camera 12, energy are installed in camera second interface simultaneously
It is enough to realize the acquisition that colour signal is carried out while highly sensitive imaging, as shown in Figure 1.
Finally it should be noted that the above content is only to illustrate the technical solution of the utility model, rather than it is practical to this
The limitation of novel protected range, those skilled in the art to the technical solution of the utility model carry out it is simple modification or
Person's equivalent replacement, all without departing from the spirit and scope of technical solutions of the utility model.
Claims (5)
1. microscopic system is just set in a kind of up-conversion luminescence near-infrared and the multi-functional wide field of colour imaging, it is characterised in that：It is described more
Microscopic system is just being set mainly by just setting fluorescence microscope mirror body, laser, collimator and extender mirror, white light source, first in function wide field
It is adjustable reflecting mirror, dichroscope, exciter filter turntable, the penetrating object lens of near-infrared, transmitting filter wheel, second adjustable
Reflecting mirror, high sensitivity detector and color camera composition；Successively collect from the bottom to top in described just setting in fluorescence microscope mirror body
At being provided with penetrating object lens of the near-infrared, exciter filter turntable, dichroscope, transmitting filter wheel, second adjustable
Reflecting mirror and high sensitivity detector, in the side just set in fluorescence microscope mirror body positioned at the described second adjustable reflecting mirror
Face is provided with the color camera, is located in the outside for just setting fluorescence microscope mirror body and installs the dichroscope position
Side is sequentially installed with the described first adjustable reflecting mirror, collimator and extender mirror and laser, and the white light source is set to described
The top of first adjustable reflecting mirror.
2. microscopic system is just set in up-conversion luminescence near-infrared according to claim 1 and the multi-functional wide field of colour imaging,
It is characterized in that：The exciter filter turntable contains exciter filter, and the transmitting filter wheel contains transmitting light optical filter,
The exciter filter and the transmitting light optical filter use short pass filter, bandpass filter or long pass filter.
3. microscopic system is just set in up-conversion luminescence near-infrared according to claim 1 or 2 and the multi-functional wide field of colour imaging,
It is characterized in that：The laser uses central wavelength for the laser of 980nm, 808nm or 730nm.
4. microscopic system is just set in up-conversion luminescence near-infrared according to claim 1 or 2 and the multi-functional wide field of colour imaging,
It is characterized in that：The white light source uses mercury lamp or LED light.
5. microscopic system is just set in up-conversion luminescence near-infrared according to claim 1 or 2 and the multi-functional wide field of colour imaging,
It is characterized in that：The high sensitivity detector is using sCMOS detector or refrigeration property EMCCD detector.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108387519A (en) * | 2018-05-03 | 2018-08-10 | 上海市质子重离子临床技术研发中心 | Microscopic system is just set in up-conversion luminescence near-infrared and the multi-functional wide field of colour imaging |
| CN109856789A (en) * | 2019-02-26 | 2019-06-07 | 中国科学院苏州生物医学工程技术研究所 | High intension super-resolution integration micro imaging system and method |
| CN110031411A (en) * | 2019-04-17 | 2019-07-19 | 南京邮电大学 | A kind of spectrum imaging system |
| CN115316961A (en) * | 2022-10-14 | 2022-11-11 | 南京诺源医疗器械有限公司 | Fluorescent image probe and handheld image detector |
-
2018
- 2018-05-03 CN CN201820646690.1U patent/CN208140563U/en active Active
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108387519A (en) * | 2018-05-03 | 2018-08-10 | 上海市质子重离子临床技术研发中心 | Microscopic system is just set in up-conversion luminescence near-infrared and the multi-functional wide field of colour imaging |
| CN109856789A (en) * | 2019-02-26 | 2019-06-07 | 中国科学院苏州生物医学工程技术研究所 | High intension super-resolution integration micro imaging system and method |
| CN109856789B (en) * | 2019-02-26 | 2021-10-22 | 中国科学院苏州生物医学工程技术研究所 | High-content super-resolution integrated microscopic imaging system and method |
| CN110031411A (en) * | 2019-04-17 | 2019-07-19 | 南京邮电大学 | A kind of spectrum imaging system |
| CN115316961A (en) * | 2022-10-14 | 2022-11-11 | 南京诺源医疗器械有限公司 | Fluorescent image probe and handheld image detector |
| CN115316961B (en) * | 2022-10-14 | 2023-02-24 | 南京诺源医疗器械有限公司 | Handheld fluorescence image probe and handheld image detector |
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| CB03 | Change of inventor or designer information |
Inventor after: Sun Jun Inventor after: Lu Jiade Inventor after: Kong Lin Inventor after: Li Yuhao Inventor after: Cheng Jingyi Inventor after: Chen Wen Inventor after: Qin Xiaojia Inventor after: Liu Jie Inventor before: Sun Jun Inventor before: Lu Jiade Inventor before: Kong Lin Inventor before: Li Yuhao Inventor before: Cheng Jingyi Inventor before: Chen Wen Inventor before: Qin Xiaojia Inventor before: Liu Jie |