CN107037048A - Imaging device, method and the imaging system of reflected signal and fluorescence signal are obtained simultaneously - Google Patents
Imaging device, method and the imaging system of reflected signal and fluorescence signal are obtained simultaneously Download PDFInfo
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- CN107037048A CN107037048A CN201610859818.8A CN201610859818A CN107037048A CN 107037048 A CN107037048 A CN 107037048A CN 201610859818 A CN201610859818 A CN 201610859818A CN 107037048 A CN107037048 A CN 107037048A
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- 238000003384 imaging method Methods 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title abstract description 15
- 238000005286 illumination Methods 0.000 claims abstract description 32
- 230000005284 excitation Effects 0.000 claims abstract description 25
- 230000005540 biological transmission Effects 0.000 claims abstract description 11
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 9
- 238000001215 fluorescent labelling Methods 0.000 claims abstract description 8
- 238000000926 separation method Methods 0.000 claims abstract description 5
- 230000003287 optical effect Effects 0.000 claims description 13
- 238000001514 detection method Methods 0.000 claims description 9
- 238000009738 saturating Methods 0.000 claims description 5
- 230000009977 dual effect Effects 0.000 description 5
- 238000004043 dyeing Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 108010022579 ATP dependent 26S protease Proteins 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000012757 fluorescence staining Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- ICIWUVCWSCSTAQ-UHFFFAOYSA-M iodate Chemical compound [O-]I(=O)=O ICIWUVCWSCSTAQ-UHFFFAOYSA-M 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 229910001507 metal halide Inorganic materials 0.000 description 1
- 150000005309 metal halides Chemical class 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000002078 nanoshell Substances 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 230000011514 reflex Effects 0.000 description 1
Classifications
-
- 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
- G01N21/84—Systems specially adapted for particular applications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/59—Transmissivity
-
- 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
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
Abstract
The present invention relates to a kind of while obtaining imaging device, method and the imaging system of reflected signal and fluorescence signal, the device includes：Illumination path, for providing collimated illumination light beam；Beam splitting device, is totally reflected for the collimated illumination light beam of the reflected signal operation wavelength to be reflected in semi-transparent semi-reflecting mode, and by the collimated illumination light beam of the fluorescence signal excitation wavelength；Object lens, for the obtained two-beam to be converged at into sample face to be imaged, reflecrtive mark reflects reflected signal, and fluorescence labeling excites and launches fluorescent emission signals；The reflected signal and fluorescent emission signals reach the beam splitting device, and reflected signal is transmitted in semi-transparent semi-reflecting mode；Fluorescent emission signals are transmitted in full impregnated mode；Light path is detected, for will be separately detected after the reflected signal of transmission and fluorescent emission signals separation.The present invention is obtained while realizing reflected signal and fluorescence signal in same system.
Description
Technical field
The present invention relates to the acquisition of information after biological specimen dyeing, in particular to acquisition reflected signal and fluorescence signal simultaneously
Imaging device, method and imaging system.
Background technology
With the development of staining technique, increasing coloring agent is applied to the research of the 26S Proteasome Structure and Function of biological specimen
In.Because different coloring agents provides different architecture cytoarchitectonics or shape information, a variety of labeling methods are often applied to
In the research of this 26S Proteasome Structure and Function, for example, form can be provided or the metal of structural information infects method and fluorescent dye method, with
And gold nano grain, nanoshell, organic dyestuff and quantum dot of molecular change information etc. can be provided.
A variety of optical image technologies have been developed at present available for the acquisition to reflected signal and fluorescence signal, but work as
Preceding method is only capable of obtaining reflected signal or fluorescence signal, for both needing to obtain reflected signal and fluorescence signal
Sample, is that, by being separately detected with different systems, and can't obtain simultaneously mostly, can only obtain respectively, for example：
Carson et al. has invented a kind of method that fluorescence signal and reflected signal can be observed in same system, and he is by cutting
Beam splitter (50/50) and dichroscope are changed, the Laser Scanning Confocal Microscope system of double mode (reflective and fluorescence type) is obtained
(Carlson A L.et al.Journal of microscopy(2007),228(1):11-24), although this mode is one
The observation to reflected signal and fluorescence signal is realized in individual system, but it needs to obtain both signals successively, not only grasps
Make inconvenience, can also expend longer time on beam splitter and dichroscope is changed, and acquisition sample signal can not be automated.
In summary, obtained while currently can not also realizing the reflected signal and fluorescence signal to biological specimen, due to
Reflected signal and fluorescence signal can provide the important composition of sample and shape information, therefore research can obtain reflected signal simultaneously
Technology with fluorescence signal is necessary.
The content of the invention
Present invention aims to overcome that above-mentioned the deficiencies in the prior art and provide a kind of while obtaining reflected signal and fluorescence
Imaging device, method and the imaging system of signal, realize it is automatic, while acquisition reflected signal and fluorescence signal.
Realizing the technical scheme that the object of the invention is used is：It is a kind of while obtaining the imaging dress of reflected signal and fluorescence signal
Put, the device includes：
Illumination path, the collimated illumination light beam for providing reflected signal operation wavelength and fluorescence signal excitation wavelength；
Beam splitting device, for the collimated illumination light beam of the reflected signal operation wavelength to be reflected in semi-transparent semi-reflecting mode,
And be totally reflected the collimated illumination light beam of the fluorescence signal excitation wavelength, the optical axis that reflection obtains two-beam keeps overlapping；
Object lens are there is provided falling to penetrate formula imaging mode, for the obtained two-beam to be converged at into sample face to be imaged, reflection
The illuminating bundle of signal operation wavelength reflects reflected signal by reflecrtive mark on face to be imaged；The photograph of fluorescence signal excitation wavelength
Fluorescence labeling on face to be imaged is excited and launches fluorescent emission signals by Mingguang City Shu Ze；The reflected signal and fluorescent emission letter
Number the beam splitting device is reached, reflected signal is transmitted in semi-transparent semi-reflecting mode；Fluorescent emission signals are transmitted in full impregnated mode；
Light path is detected, for will be separately detected after the reflected signal of transmission and fluorescent emission signals separation.
In addition, the present invention also provides a kind of imaging for obtaining reflected signal and fluorescence signal simultaneously by above-mentioned imaging device
Method, this method includes：
Illumination path sends the collimated illumination light beam of reflected signal operation wavelength and fluorescence signal excitation wavelength, and by light beam
It is oblique to be mapped on the beam splitting device with three wavelength operation windows；
Beam splitting device reflects the collimated illumination light beam of the reflected signal operation wavelength in semi-transparent semi-reflecting mode, and will
The collimated illumination light beam total reflection of the fluorescence signal excitation wavelength, the optical axis that reflection obtains two-beam keeps overlapping；
The obtained two-beam is converged at sample face to be imaged, the illuminating bundle quilt of reflected signal operation wavelength by object lens
Reflecrtive mark reflects reflected signal on face to be imaged；The illuminating bundle of fluorescence signal excitation wavelength is then by fluorescence on face to be imaged
Mark excites and launches fluorescent emission signals；The reflected signal and fluorescent emission signals reach the beam splitting device, reflection
Signal is transmitted in semi-transparent semi-reflecting mode；Fluorescent emission signals are transmitted in full impregnated mode；
Detection light path will be separately detected after the reflected signal of transmission and fluorescent emission signals separation.
Present invention also offers imaging system, the imaging system includes above-mentioned while obtaining reflected signal and fluorescence signal
Imaging device.
The present invention has following advantages compared with prior art：
1st, obtained while realizing reflected signal and fluorescence signal in same system；
2nd, the automation for realizing reflected signal and fluorescence signal is obtained, and filter set is changed without artificial.
Brief description of the drawings
Fig. 1 obtains the image device structure schematic diagram of reflected signal and fluorescence signal for the present invention simultaneously.
Fig. 2 is the working method schematic diagram of the beam splitting device with three wavelength operation windows in the present invention.
Fig. 3 is a kind of structural representation of preferred embodiment of imaging device of the present invention.
Fig. 4 a are the result figure that Fig. 3 equipment therefors obtain Gorky's staining signals (reflected signal) simultaneously；Fig. 4 b are iodate
The result figure of third pyridine staining signals (fluorescence signal).
Embodiment
The present invention is described in further detail with specific embodiment below in conjunction with the accompanying drawings.
As shown in figure 1, the present embodiment obtains reflected signal simultaneously and the imaging device of fluorescence signal is illuminated including dual wavelength
Light path 1, the beam splitting device 2 with three wavelength operation windows, object lens 3, sample 4 and binary channels detection light path 5.By dual wavelength
Light beam after the collimated illumination of illumination path 1 is oblique to be mapped on the beam splitting device 2 with three wavelength operation windows, in the present embodiment,
Dual wavelength illumination path 1 provides the illuminating bundle of reflected signal operation wavelength and the illuminating bundle of fluorescence signal excitation wavelength, its
In, the illuminating bundle of reflected signal operation wavelength is by the beam splitting device 2 with three wavelength operation windows in semi-transparent semi-reflecting mode
Reflection, the illuminating bundle of the excitation wavelength of fluorescence signal is totally reflected by the beam splitting device 2 with three wavelength operation windows, is reflected
Two-beam optical axis keeps overlapping afterwards, incides the rear cylinder of object lens 3.Then object lens 3 converge at two beam illuminating bundles after reflection
The face to be imaged of sample 4, wherein, the illuminating bundle of reflected signal operation wavelength is reflected by reflecrtive mark on face to be imaged to be obtained
Reflected signal；Fluorescence labeling on face to be imaged is then excited and launches fluorescence hair by the illuminating bundle of the excitation wavelength of fluorescence signal
Penetrate signal.Reflected signal and fluorescent emission signals optical axis coincidence, return light path after being assembled through object lens 3, reaching has three wavelength
The beam splitting device 2 of operation window.Reflected signal is saturating in semi-transparent semi-reflecting mode by the beam splitting device 2 with three wavelength operation windows
Penetrate；Fluorescent emission signals are transmitted by the beam splitting device 2 with three wavelength operation windows in full impregnated mode.Enter bilateral after transmission
Road detects light path 5.Then, binary channels detection light path 5 is separately detected after reflected signal is separated with fluorescent emission signals, and by institute
The signal of detection is delivered to computer, can obtain the reflected image and fluoroscopic image in face to be imaged after being rebuild through algorithm respectively, and
And the reflection obtained and fluoroscopic image can be with spatial registrations.
The architectural feature that the present invention has the beam splitting device 2 of three wavelength operation windows provides for first wave length operation window
Semi-transparent semi-reflecting, the reflection of second wave length operation window offer fluorescence signal excitation wavelength, the 3rd wavelength working window of reflected signal
Mouth provides the transmission of fluorescent emission signals launch wavelength.The wavelength of reflected signal is close with the excitation wavelength of fluorescence signal, instead
Penetrate the wavelength of signal and the wavelength of transmitted light spectrum of fluorescent emission signals has an obvious boundary, and it is the wavelength of reflected signal, glimmering
There is no overlapping region or only between the excitation wavelength of optical signal and the wavelength of transmitted light of fluorescent emission signals these three wavelength
There is a small amount of overlapping region.The present embodiment has the working method of the beam splitting device 2 of three wavelength operation windows as shown in Fig. 2 its
In, label 6,7,8 corresponds to three wavelength operation windows of this beam splitting device 2 respectively.Operation window 6 is semi-transparent for this beam splitting device
Half anti-operation window, reflected signal is just by operation window 6 with semi-transparent semi-reflecting mode incidence and transmission；Operation window 7 is this point
The total reflection operation window of beam device, excitation wavelength incidence in the way of total reflection by operation window 7 of fluorescence signal；Working window
Mouth 8 is the total transmissivity operation window of this beam splitting device, and fluorescence signal is transmitted by operation window 8 in total transmissivity mode.
Fig. 3 is a kind of structural representation for preferred embodiment that apparatus of the present invention are applied in Structured Illumination micro-imaging
Figure, dual wavelength illumination path 1 used in the present embodiment includes white light source 9, collimation lens 10, spatial light modulator 11, speculum
12nd, tube lens 13 and low pass filters 14, the formation of dual wavelength illumination path 1 reflected signal operation wavelength used in the present embodiment
The process of illuminating bundle and the illuminating bundle of fluorescence signal excitation wavelength is：The light that white light source 9 is sent passes through collimation lens 10
After be radiated in spatial light modulator 11, reflex to mirror by speculum 12 by the modulated structure light of spatial light modulator 11
Cylinder lens 13 on, formed directional light by low pass filters 14 retread 45 degree incidence three wavelength operation window beam splitting devices 15.
The illuminating bundle of reflected signal operation wavelength is by the beam splitting device 15 with three wavelength operation windows with semi-transparent half
Antimode reflects；The illuminating bundle of the excitation wavelength of fluorescence signal is all-trans by the beam splitting device 15 with three wavelength operation windows
Penetrate；Two-beam optical axis keeps overlapping after reflection, impinges perpendicularly on the rear cylinder of object lens 16.Then, object lens 16 assemble illuminating bundle
In the face to be imaged of sample 17.The illuminating bundle of reflected signal operation wavelength is reflected by reflecrtive mark on face to be imaged obtains anti-
Penetrate signal；Fluorescence labeling on face to be imaged is then excited and launches fluorescent emission by the illuminating bundle of the excitation wavelength of fluorescence signal
Signal.Reflected signal and fluorescent emission signals optical axis coincidence, return light path after being assembled through object lens 16, reaching has three wavelength works
Make the beam splitting device 15 of window.Reflected signal is saturating in semi-transparent semi-reflecting mode by the beam splitting device 15 with three wavelength operation windows
Penetrate；Fluorescent emission signals are transmitted by the beam splitting device 15 with three wavelength operation windows in full impregnated mode.Two beams after transmission
Light enters dichroscope 19 after the convergence of tube lens 18, and reflected signal is separated with fluorescent emission signals, fluorescent emission
Signal is then filtered by optical filter 21, then respectively by the detection of detector 20 and 22.Then, the signal of detection is transported to calculating
Machine, obtains the reflected image and fluoroscopic image in face to be imaged, and the reflection obtained and fluoroscopic image respectively after being rebuild through algorithm
Can be with spatial registration.
Wherein, the parameter of each part used is specially in the present embodiment：Light source 9 is using the production of Lumen Dynamics companies
X-cite exact metal halide lights；Spatial light modulator 11 uses specification for 0.7XGA digital micromirror array；Filter
Mating plate 14,21 is that the model of Semrock companies production is respectively BSP01-532R low pass filter, FF01-665_150 height
Pass filter, beam splitting device 15, dichroscope 19 with three wavelength operation windows are the model difference that Semrock companies produce
For FF560-Di01, FF562-Di03 saturating short anti-dichroscope of length；The object lens 16 of imaging are Olympus companies of Japan
NA1.0/20 × achromatic objective；Detector 21 and 23 is the sCMOS cameras that Hamamatsu companies of Japan produce, pixel specification
For 2048 × 2048.
The Gorky's dyeing reflected signal and propidium iodide stain for obtaining Mice brain tissues simultaneously using Fig. 3 shown devices are glimmering
Respectively as shown in figures 4 a and 4b, Fig. 4 a are the Gorky's dyeing reflected signal result figure detected, Fig. 4 b to the result of optical signal
For propidium iodide fluorescence staining signals result figure.
Obtain a kind of application of the imaging device of reflected signal and fluorescence signal, the imaging device energy simultaneously as the present invention
It is enough in co-focusing imaging, Structured Illumination imaging or the imaging of other Optics in Microscope of broadband light or the illumination of multi beam narrow-band spectrum
Imaging system.When in the incident light of imaging system simultaneously comprising fluorescent exciting and reflected light incident light, using possessing this hair
It can be achieved to obtain reflected signal and fluorescence signal simultaneously after bright imaging device.
Claims (10)
1. it is a kind of while obtaining the imaging device of reflected signal and fluorescence signal, it is characterised in that including：
Beam splitting device with three wavelength operation windows, for reflecrtive mark on face to be imaged to be reflected into reflected signal with half
The transmission of saturating half antimode, and fluorescence labeling on face to be imaged is excited and to launch fluorescent emission signals saturating in full impregnated mode
Penetrate；
Light path is detected, for will be separately detected after the reflected signal of transmission and fluorescent emission signals separation.
2. obtain the imaging device of reflected signal and fluorescence signal simultaneously according to claim 1, it is characterised in that also wrap
Include：
Illumination path, the collimated illumination light beam for providing reflected signal operation wavelength and fluorescence signal excitation wavelength；Described point
Beam device reflects the collimated illumination light beam of the reflected signal operation wavelength in semi-transparent semi-reflecting mode, and the fluorescence is believed
The collimated illumination light beam total reflection of number excitation wavelength, the optical axis for respectively obtaining two-beam keeps overlapping.
Object lens, for two light beam to be converged at into sample face to be imaged, the illuminating bundle of reflected signal operation wavelength is treated into
Reflecrtive mark reflects reflected signal in image planes；The illuminating bundle of fluorescence signal excitation wavelength is then by fluorescence labeling on face to be imaged
Excite and launch fluorescent emission signals；The reflected signal and fluorescent emission signals reach the beam splitting device, reflected signal
Transmitted in semi-transparent semi-reflecting mode, fluorescent emission signals are transmitted in full impregnated mode.
3. obtain the imaging device of reflected signal and fluorescence signal simultaneously according to claim 2, it is characterised in that also wrap
Include：
Computer, for receiving reflected signal and the fluorescent emission signals that the detection light path is detected, calculating obtains to be imaged
The reflected image and fluoroscopic image in face.
4. obtaining the imaging device of reflected signal and fluorescence signal simultaneously according to any one of claims 1 to 3, its feature exists
In：Three operation windows of the beam splitting device just include the exciting light of fluorescence signal and the incident light of reflected signal, fluorescence letter
Number transmitting light and reflected signal emergent light.
5. it is a kind of while obtaining the imaging method of reflected signal and fluorescence signal, it is characterised in that including：
The illuminating bundle of reflected signal operation wavelength reflects reflected signal by reflecrtive mark on face to be imaged；Fluorescence signal is excited
Fluorescence labeling on face to be imaged is then excited and launches fluorescent emission signals by the illuminating bundle of wavelength；The reflected signal and glimmering
Light emission signal reaches the beam splitting device, and reflected signal is transmitted in semi-transparent semi-reflecting mode；Fluorescent emission signals are in full impregnated mode
Transmission；
Detection light path will be separately detected after the reflected signal of transmission and fluorescent emission signals separation.
6. obtain the imaging method of reflected signal and fluorescence signal simultaneously according to claim 5, it is characterised in that including：
Illumination path sends the collimated illumination light beam of reflected signal operation wavelength and fluorescence signal excitation wavelength, and light beam is oblique
It is mapped on the beam splitting device with three wavelength operation windows；
Beam splitting device with three wavelength operation windows is by the collimated illumination light beam of the reflected signal operation wavelength with semi-transparent
Half antimode is reflected, and the collimated illumination light beam of the fluorescence signal excitation wavelength is totally reflected, and reflection obtains two-beam
Optical axis keeps overlapping.
7. obtain the imaging method of reflected signal and fluorescence signal simultaneously according to claim 6, it is characterised in that：Described three
The beam splitting device of individual wavelength operation window is that shorter wavelength is reflected and longer wavelength is transmitted, or to shorter
Wavelength is transmitted and longer wavelength is reflected.
8. a kind of imaging system, it is characterised in that：Including obtained simultaneously described in claim 1 reflected signal and fluorescence signal into
As device.
9. imaging system according to claim 1, it is characterised in that also include：
Illumination path, the collimated illumination light beam for providing reflected signal operation wavelength and fluorescence signal excitation wavelength；Described point
Beam device reflects the collimated illumination light beam of the reflected signal operation wavelength in semi-transparent semi-reflecting mode, and the fluorescence is believed
The collimated illumination light beam total reflection of number excitation wavelength, the optical axis for respectively obtaining two-beam keeps overlapping.
Object lens, for two light beam to be converged at into sample face to be imaged, the illuminating bundle of reflected signal operation wavelength is treated into
Reflecrtive mark reflects reflected signal in image planes；The illuminating bundle of fluorescence signal excitation wavelength is then by fluorescence labeling on face to be imaged
Excite and launch fluorescent emission signals；The reflected signal and fluorescent emission signals reach the beam splitting device, reflected signal
Transmitted in semi-transparent semi-reflecting mode, fluorescent emission signals are transmitted in full impregnated mode.
10. imaging system according to claim 9, it is characterised in that also include：
Computer, for receiving reflected signal and the fluorescent emission signals that the detection light path is detected, calculating obtains to be imaged
The reflected image and fluoroscopic image in face.
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| CN108836262A (en) * | 2018-04-11 | 2018-11-20 | 秦少平 | A kind of induced fluorescence spectrum picture fusion evaluation optical path |
| CN109288489A (en) * | 2018-10-17 | 2019-02-01 | 中国科学院苏州生物医学工程技术研究所 | A kind of imaging scope |
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| CN108836262A (en) * | 2018-04-11 | 2018-11-20 | 秦少平 | A kind of induced fluorescence spectrum picture fusion evaluation optical path |
| CN108836262B (en) * | 2018-04-11 | 2021-08-31 | 秦少平 | Induced fluorescence spectrum image fusion image light path |
| CN109288489A (en) * | 2018-10-17 | 2019-02-01 | 中国科学院苏州生物医学工程技术研究所 | A kind of imaging scope |
| CN109288490A (en) * | 2018-10-17 | 2019-02-01 | 中国科学院苏州生物医学工程技术研究所 | A kind of imaging scope |
| CN113495074A (en) * | 2020-04-07 | 2021-10-12 | 大量科技股份有限公司 | Visual inspection system |
| CN113640219A (en) * | 2021-07-13 | 2021-11-12 | 中国科学院半导体研究所 | Linkage switching device for excitation light, beam splitter and optical filter of spectrometer |
| CN113640219B (en) * | 2021-07-13 | 2024-02-27 | 中国科学院半导体研究所 | Linkage switching device for excitation light, beam splitter and optical filter of spectrometer |
| CN113502207A (en) * | 2021-08-18 | 2021-10-15 | 长春长光辰英生物科学仪器有限公司 | Multifunctional cell sorting device based on laser system and operation method |
| CN113502207B (en) * | 2021-08-18 | 2022-11-15 | 长春长光辰英生物科学仪器有限公司 | Multifunctional cell sorting device based on laser system and operation method |
| CN114134025A (en) * | 2021-11-19 | 2022-03-04 | 中国科学院长春光学精密机械与物理研究所 | Gene sequencing system and sequencing method thereof |
| CN114134025B (en) * | 2021-11-19 | 2023-08-22 | 中国科学院长春光学精密机械与物理研究所 | Gene sequencing system and sequencing method thereof |
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