CN208736797U - The Raman scattering Quick Acquisition and imaging device of fixed wave length - Google Patents
The Raman scattering Quick Acquisition and imaging device of fixed wave length Download PDFInfo
- Publication number
- CN208736797U CN208736797U CN201821011459.1U CN201821011459U CN208736797U CN 208736797 U CN208736797 U CN 208736797U CN 201821011459 U CN201821011459 U CN 201821011459U CN 208736797 U CN208736797 U CN 208736797U
- Authority
- CN
- China
- Prior art keywords
- laser
- raman
- raman scattering
- signal
- filter
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000001069 Raman spectroscopy Methods 0.000 title claims abstract description 225
- 238000003384 imaging method Methods 0.000 title claims abstract description 33
- 230000003287 optical effect Effects 0.000 claims abstract description 66
- 238000001914 filtration Methods 0.000 claims abstract description 7
- 230000005284 excitation Effects 0.000 claims description 17
- 238000002834 transmittance Methods 0.000 claims description 13
- 238000007334 copolymerization reaction Methods 0.000 claims description 8
- 230000009471 action Effects 0.000 claims description 3
- 230000005622 photoelectricity Effects 0.000 claims 1
- 239000011521 glass Substances 0.000 description 13
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical group [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 description 8
- 229910052805 deuterium Inorganic materials 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 7
- 238000001237 Raman spectrum Methods 0.000 description 7
- 230000005540 biological transmission Effects 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 230000013011 mating Effects 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 2
- 239000005383 fluoride glass Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241001360526 Escherichia coli ATCC 25922 Species 0.000 description 1
- YZCKVEUIGOORGS-IGMARMGPSA-N Protium Chemical compound [1H] YZCKVEUIGOORGS-IGMARMGPSA-N 0.000 description 1
- 241000720974 Protium Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 125000001967 indiganyl group Chemical group [H][In]([H])[*] 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The utility model relates to the Raman scattering Quick Acquisitions and imaging device of a kind of fixed wave length, including laser emission element, reflection microscope group, Raman scattering signal filter element and Raman scattering signal acquisition unit.Laser emission element is for obtaining the laser after power adjustment, microscope group is reflected to be used for the laser reflection after power adjustment to sample surfaces, Raman scattering signal filter element is used to Raman scattering signal being divided into echo signal and two strands of background noise, and respectively reduce the wave-length coverage of two strands of signals, Raman scattering signal acquisition unit is for collecting two strands of Raman signals.Compared with prior art, the utility model saves grating beam splitting, uses optical filter Filtration Goal wavelength signals range instead, and use photon counting or photomultiplier tube collecting signal, greatly improves the Raman signal intensity of sample, reduce acquisition time, realizes fast imaging.Optical components, the costs such as grating is saved substantially to lower than existing Raman product in the market.
Description
Technical field
The utility model relates to a kind of Raman image equipment, the Raman scattering more particularly, to a kind of fixed wave length is quickly adopted
Collection and imaging device.
Background technique
Chinese patent CN107462566A discloses the Raman spectrometer for detecting specific narrow wave-number range, the Raman light
Spectrometer includes laser transmitting set, spectra collection equipment, data processing equipment, and the laser transmitting set is for obtaining power tune
Excitation beam after whole, the spectra collection equipment are used to that the excitation beam after power adjustment to be made to get to sample surfaces generation Raman
Scattering, and orientation Rayleigh scattering light is filtered off, orientation Raman diffused light is obtained, and be received by the receiver orientation Raman scattering, institute
Stating data processing equipment is the computer equipment connecting with receiver, formation, analysis and judgement for obtained Raman spectrum；It is described
Receiver be single-photon detector, orientation Raman diffused light finally received by single-photon detector.
As above-mentioned publication technology, in existing qualitative or quantitative Raman spectroscopy, usually using a certain
The laser excitation sample of wavelength, after Raman scattering signal passes through grating beam splitting, according to Wavelength distribution by detector (usually CCD
Or EMCCD) read, to generate Raman spectrum.Existing Raman spectroscopy can completely react sample in different wave length
Raman signal intensity distribution, but disadvantage is to weaken with grating beam splitting bring Raman signal, and what is generated therewith adopt
Spectrum and image taking speed slow down.
Utility model content
The purpose of this utility model is exactly to provide a kind of fixed wave length to overcome the problems of the above-mentioned prior art
Raman scattering Quick Acquisition and imaging device.
The purpose of this utility model can be achieved through the following technical solutions:
A kind of the Raman scattering Quick Acquisition and imaging device of fixed wave length, comprising:
Raman scattering signal filter element: for filtering the Raman scattering signal that sample generates under laser action by wavelength
The filtering of device or equipment selectivity,
Raman scattering signal acquisition unit: for receiving the Raman scattering signal for passing through Raman scattering signal filter element,
Use photon reading device or equipment as detector.
Especially, it should be noted that the Raman scattering Quick Acquisition and imaging device of the utility model fixed wave length do not use
Grating.
In an embodiment of the utility model, wavelength filter device used in Raman scattering signal filter element or
Equipment is one or more optical filters.
In an embodiment of the utility model, the photon reading device or equipment of Raman scattering signal acquisition unit
Detector be one or more photon counters or photomultiplier tube.
In an embodiment of the utility model, the Raman scattering signal filter element includes beam splitting chip, first
Narrow band filter, the second narrow band filter, it is narrow through first after the beam splitting chip is used to that a part of Raman signal to be made to penetrate beam splitting chip
Band optical filter filters；After remaining Raman signal is reflected by beam splitting chip, filtered after the second narrow band filter.
In an embodiment of the utility model, the Raman scattering signal filtered after the first narrow band filter is by
One detector is read, and the Raman scattering signal filtered after the second narrow band filter is read by the second detector.
In an embodiment of the utility model, the beam splitting chip is a:b beam splitting chip, and a and b are respectively light transmission
The ratio of light intensity after crossing beam splitting chip and being reflected by beam splitting chip, a:b is greater than 0, less than 1.
For example, the beam splitting chip is 10:90 beam splitting chip, after 90% Raman signal penetrates beam splitting chip, through the first narrow-band-filter
It is read after piece by the first detector, after 10% Raman signal is reflected by beam splitting chip, is visited after the second narrow band filter by second
Device is surveyed to read.
In an embodiment of the utility model, the beam splitting chip working range is at least covered than laser excitation wave
Long longer any 100cm-1The continuous wavelength range intervals of wave number；
Preferably, the beam splitting chip working range is at least covered than laser excitation wavelength red shift 2000cm-1It arrives
2300cm-1The wavelength of wave-number range.
In an embodiment of the utility model, first narrow band filter is selected red in laser excitation wavelength
Move direction 2040cm-1To 2300cm-1Optical transmittance in wave-number range is greater than 93% optical filter.
In an embodiment of the utility model, second narrow band filter is selected red in laser excitation wavelength
Move direction 1800cm-1-2040cm-1Or 2260cm-1To 2700cm-1The 30cm of middle arbitrary continuation in wave-number range-1Or 30cm-1
Optical transmittance in above wave-number range is greater than 93% optical filter.
Such as first narrow band filter selects 600/14nm to read the Raman scattering signal within the scope of 588-608nm
With the region Raman background signal；
Such as second narrow band filter selects 591/6nm to read the Raman background signal within the scope of 586-596nm.
It further include laser emission element, laser emission element is for generating in an embodiment of the utility model
Laser and generation Raman scattering, the laser emission element include laser, laser narrow-band optical filter, and the laser is for producing
Raw laser, the laser narrow-band optical filter are used to filter out the miscellaneous line in laser.
In an embodiment of the utility model, the laser emission element further includes laser power attenuator, is used
In carrying out different degrees of decaying to laser intensity, to adjust laser intensity.
In an embodiment of the utility model, the laser can be 488nm laser, 514nm laser
One of device, 532nm laser, 633nm laser or 785nm laser.
In an embodiment of the utility model, the laser power attenuator selects 6 groups of attenuators with strong to laser
Degree carries out different degrees of decaying, to better meet protection sample and enhance the function of Raman signal.
In a specific embodiment of the utility model, the laser selects the 532nm laser of power adjustable, institute
Stating laser narrow-band optical filter selects transmitance at 532nm to be greater than 90%, transmittance curve full width at half maximum 2nm, to filter out in laser
Miscellaneous line.
In a specific embodiment of the utility model, the Raman scattering signal acquisition unit further includes Raman filter
Mating plate and object lens, the Raman optical filter, which is used to reflect after the laser reflection that microscope group launches, focuses to sample by object lens
On, the object lens are used to collect the Raman scattering signal and Rayleigh scattering signal of sample scattering, and make Raman scattering signal and auspicious
Sharp scattered signal returns to Raman optical filter；
The Rayleigh scattering signal that the Raman optical filter is also used to that sample is stopped to scatter, through the Raman scattering of sample scattering
Signal.
It further include the glass slide for placing sample in a specific embodiment of the utility model, the load glass
Piece is located on auto-translating platform.The glass slide is advisable with can more preferably reduce the material of laser reflection or structure.
It further include reflection microscope group in a specific embodiment of the utility model, the reflection microscope group includes first
Reflecting mirror and the second reflecting mirror.
In a specific embodiment of the utility model, the Raman scattering signal acquisition unit further includes that copolymerization is burnt
Pin hole, slit, the burnt pin hole of the copolymerization and slit are arranged at Raman optical filter rear, for projecting beam splitting after passing through laser
On piece, the burnt pin hole of the copolymerization promote sample signal intensity under non-confocal mode for stopping spurious signal under confocal mode.
The working principle of the utility model is: after the Raman scattering of laser excitation sample, by by specific optical filterSelection PropertyThe Raman photon through particular range of wavelengths, and using photon reading device or equipment capture and read Raman signal it is strong
Degree.To realize the purpose of fast Raman detection and Raman image.
Such as: usually in H2In the environment of O, c h bond, the corresponding table in Raman spectrum are generated in cell normal metabolic processes
It is now 2800-3000cm-1C-H Raman signal (Fig. 2).After a part of protium in cell culture environment is substituted by deuterium,
Cell generates C-D key in the metabolic process, and correspondence shows as 1800-2800cm in Raman spectrum-1C-D Raman signal (figure
2).The utility model is the technology of the intensity of C-D signal in quick detection cell.
The Raman fast imaging techniques of fixed wave length are one kind, based on detection and the closely related C-D of cell metabolic activity
The variation of peak raman scattering intensity, and reach the technological means of the judgement cell activity of fast accurate, cell susceptibility degree.Since Raman is believed
Number wavelength value changes with excitation light wave long value, therefore needs to calculate the wavelength at carbon deuterium peak according to formula. Wavenumber(cm-1)=
(1/λ0-1/λ1)*107
λ in the formula0Excitation wavelength, λ1Raman signal wavelength
Compared with prior art, the utility model have the following advantages that and the utility model has the advantages that
1, cell fast imaging: saving grating beam splitting, uses optical filter instead and chooses target wavelength signal-obtaining, greatly improves
The Raman signal intensity of sample reduces acquisition time, realizes fast imaging.
2, fast Raman detects.Such as cell characteristic rapid screening: being believed by the Raman of quick detection cell C-D key
Number intensity, quickly identification and screening cell feature related to this.
3, Raman fluidic cell sorts: quickly reading cell Raman signal, real Fast synchronization cell sorting.
4, optical components, the costs such as grating is saved substantially to lower than existing Raman product in the market.
Detailed description of the invention
Fig. 1 is the Raman scattering Quick Acquisition and imaging device structure schematic diagram of fixed wave length in embodiment 1；
Label in Fig. 1: 1, laser, 2, laser narrow-band optical filter, 3, laser power attenuator, the 4, first reflecting mirror, 5,
Second reflecting mirror, 6, Raman optical filter, 7, object lens, 8, glass slide, 9, auto-translating platform, 10, the burnt pin hole of copolymerization, 11, slit,
12, beam splitting chip, the 13, first narrow band filter, the 14, first detector, the 15, second narrow band filter, the 16, second detector.
Fig. 2 is to absorb the single cell Raman spectrum of the Escherichia coli of different degrees of deuterium.
Fig. 3 is that grating beam splitting is changed to optical filter in the fixed wave length fast Raman Signal Collection Technology of the utility model
Afterwards, the unicellular Raman signal of the Escherichia coli of deuterium is absorbed.
Specific embodiment
A kind of the Raman scattering Quick Acquisition and imaging device of fixed wave length, comprising:
Raman scattering signal filter element: for filtering the Raman scattering signal that sample generates under laser action by wavelength
The filtering of device or equipment selectivity,
Raman scattering signal acquisition unit: for receiving the Raman scattering signal for passing through Raman scattering signal filter element,
Use photon reading device or equipment as detector.
Especially, it should be noted that the Raman scattering Quick Acquisition and imaging device of fixed wave length do not use grating.
In one embodiment, wavelength filter device or equipment used in Raman scattering signal filter element be one or
Multiple optical filters.
In one embodiment, the photon reading device of Raman scattering signal acquisition unit or the detector of equipment are one
A or multiple photon counters or photomultiplier tube.
In one embodiment, the Raman scattering signal filter element includes beam splitting chip, the first narrow band filter,
Two narrow band filters, the beam splitting chip through the first narrow band filter for filtering after so that a part of Raman signal is penetrated beam splitting chip；
After remaining Raman signal is reflected by beam splitting chip, filtered after the second narrow band filter.
In one embodiment, the Raman scattering signal filtered after the first narrow band filter is read by the first detector
It takes, the Raman scattering signal filtered after the second narrow band filter is read by the second detector.
In one embodiment, the beam splitting chip is a:b beam splitting chip, a and b be respectively light transmitted through beam splitting chip and by
The ratio of light intensity after beam splitting chip reflection, a:b is greater than 0, less than 1.
For example, the beam splitting chip is 10:90 beam splitting chip, after 90% Raman signal penetrates beam splitting chip, through the first narrow-band-filter
It is read after piece by the first detector, after 10% Raman signal is reflected by beam splitting chip, is visited after the second narrow band filter by second
Device is surveyed to read.
In one embodiment, the beam splitting chip working range at least covers longer than laser excitation wavelength any
100cm-1The continuous wavelength range intervals of wave number；Preferably, the beam splitting chip working range is at least covered than laser excitation wave
Long red shift 2000cm-1To 2300cm-1The wavelength of wave-number range.
In one embodiment, first narrow band filter is selected in laser excitation wavelength red shift direction
2040cm-1To 2300cm-1Optical transmittance in wave-number range is greater than 93% optical filter.
In one embodiment, second narrow band filter is selected in laser excitation wavelength red shift direction
1800cm-1-2040cm-1Or 2260cm-1To 2700cm-1The 30cm of middle arbitrary continuation in wave-number range-1Or 30cm-1Above
Optical transmittance in wave-number range is greater than 93% optical filter.
Such as first narrow band filter selects 600/14nm to read the Raman scattering signal within the scope of 588-608nm
With the region Raman background signal；
Such as second narrow band filter selects 591/6nm to read the Raman background signal within the scope of 586-596nm.
It in one embodiment, further include laser emission element, laser emission element is for generating laser and generating drawing
Graceful scattering, the laser emission element include laser, laser narrow-band optical filter, and the laser is described for generating laser
Laser narrow-band optical filter is used to filter out the miscellaneous line in laser.
In one embodiment, the laser emission element further includes laser power attenuator, for laser intensity
Different degrees of decaying is carried out, to adjust laser intensity.
In one embodiment, the laser can be 488nm laser, 514nm laser, 532nm laser
One of device, 633nm laser or 785nm laser.
In one embodiment, the laser power attenuator selects 6 groups of attenuators to carry out different journeys to laser intensity
The decaying of degree, to better meet protection sample and enhance the function of Raman signal.
In a specific embodiment, the laser selects the 532nm laser of power adjustable, the laser narrow-band filter
Mating plate selects transmitance at 532nm to be greater than 90%, transmittance curve full width at half maximum 2nm, to filter out the miscellaneous line in laser.
In a specific embodiment, the Raman scattering signal acquisition unit further includes Raman optical filter and object lens,
The Raman optical filter is used to reflect after the laser reflection that microscope group launches and focuses to sample by object lens, and the object lens are used
In the Raman scattering signal and Rayleigh scattering signal of collection sample scattering, and return to Raman scattering signal and Rayleigh scattering signal
Raman optical filter；
The Rayleigh scattering signal that the Raman optical filter is also used to that sample is stopped to scatter, through the Raman scattering of sample scattering
Signal.
It in a specific embodiment, further include glass slide for placing sample, the glass slide is located at automatic flat
In moving stage.The glass slide is advisable with can more preferably reduce the material of laser reflection or structure.
It in a specific embodiment, further include reflection microscope group, the reflection microscope group includes the first reflecting mirror and second
Reflecting mirror.
In a specific embodiment, the Raman scattering signal acquisition unit further includes being copolymerized burnt pin hole, slit, institute
The burnt pin hole of copolymerization and slit setting are stated at Raman optical filter rear, it is described total for projecting beam splitting chip after passing through laser
Pin hole is focused for stopping spurious signal under confocal mode, promotes sample signal intensity under non-confocal mode.
The utility model also provides Raman scattering Quick Acquisition and the application of imaging device of the fixed wave length, and Raman dissipates
It penetrates signal to project on beam splitting chip, a part of Raman signal is visited after the first narrow band filter by first again by beam splitting chip herein
It surveys device to read, the Raman signal of another part is read after the second narrow band filter by the second detector again by beam splitting chip；The
One detector reads the Raman scattering signal and part background signal of specific sections, and the second detector reads the background of Raman scattering
Signal；Finally by algorithm, the background signal that the first detector is read is deducted, to obtain the target Raman signal of specific sections.
Specifically, exciting light is issued through laser in experimentation, miscellaneous line is filtered out by laser narrow-band optical filter, then through laser function
After rate attenuator adjusting strength, the incident angle after being optimized laser by reflection microscope group is projected to Raman optical filter；Laser warp
It is focused on the sample on glass slide after the reflection of Raman optical filter by object lens；Auto-translating platform can move automatically according to software set
Dynamic sample position, to reach scanning imagery purpose；It is received by the Raman scattering signal and Rayleigh scattering signal of sample scattering by object lens
Return Raman optical filter after collection, is blocked in this Rayleigh scattering signal and Raman scattering signal penetrates Raman optical filter；For example, drawing
Graceful scattered signal using being copolymerized burnt pin hole and slit projects on beam splitting chip, this 90% Raman signal by beam splitting chip again
It is read after the first narrow band filter by the first detector, 10% Raman signal is by beam splitting chip again through the second narrow band filter
(Raman background signal area of the transmission range before C-D wavelength region, and C-D Raman wavelength region transmitance is 0) quilt afterwards
Second detector is read；First detector reads C-D Raman signal and Raman background signal, the only drawing that the second detector is read
Graceful background signal；Finally by algorithm, the background signal that the first detector is read is deducted, to obtain pure C-D Raman signal.
The utility model is described in detail in the following with reference to the drawings and specific embodiments.
Embodiment
With reference to Fig. 1, a kind of the Raman scattering Quick Acquisition and imaging device of fixed wave length, including it is laser emission element, anti-
Penetrate microscope group, Raman scattering signal filter element and Raman scattering signal acquisition unit.Laser emission element is for obtaining power tune
Laser after whole, reflection microscope group are used for the laser reflection after power adjustment to Raman scattering signal filter element, Raman scattering
Signal filter element is used to that the laser after power adjustment to be made to get to sample surfaces generation Raman scattering, and it is auspicious to stop sample to scatter
Sharp scattered signal, through the Raman scattering signal of sample scattering, Raman scattering signal acquisition unit is for receiving Raman scattering letter
Number, Raman scattering signal acquisition unit includes beam splitting chip 12, and beam splitting chip 12 is used to for Raman scattering signal being divided into two strands, and respectively
It is read by two groups of detectors.
Specifically, laser emission element includes laser 1, laser narrow-band optical filter 2 and laser power attenuator 3, swash
Light device 1 is used to filter out the miscellaneous line in laser for generating laser, laser narrow-band optical filter 2, and laser power attenuator 3 is used for sharp
Luminous intensity carries out different degrees of decaying, to adjust laser intensity.Laser power attenuator 3 selects 6 groups of attenuators with strong to laser
Degree carries out different degrees of decaying, to better meet protection sample and enhance the function of Raman signal.
Laser 1 usually the 532nm laser of optional power adjustable or the laser of other optional wavelength, including 488nm,
514nm, 633nm, 785nm etc.；For match 532nm laser, laser narrow-band optical filter 2 usually optional 532/2nm, i.e.,
Transmitance is greater than 90%, transmittance curve full width at half maximum 2nm at 532nm, to filter out the miscellaneous line in laser.
Raman scattering signal filter element includes Raman optical filter 6 and object lens 7, and Raman optical filter 6 will be for that will reflect microscope group
It is focused on sample after laser reflection after the power adjustment launched by object lens 7, object lens 7 are used to collect the drawing of sample scattering
Graceful scattered signal and Rayleigh scattering signal, and Raman scattering signal and Rayleigh scattering signal is made to return to Raman optical filter 6, Raman filter
The Rayleigh scattering signal that mating plate 6 is also used to that sample is stopped to scatter, through the Raman scattering signal of sample scattering.Including for placing
The glass slide 8 of sample, glass slide 8 are located on auto-translating platform 9.Glass slide is can more preferably reduce the material or knot of laser reflection
Structure is advisable.
Reflecting microscope group includes the first reflecting mirror 4 and the second reflecting mirror 5.
Raman scattering signal acquisition unit includes being copolymerized burnt pin hole 10, slit 11, beam splitting chip 12, the first narrow band filter
13, the first detector 14, the second narrow band filter 15 and the second detector 16 are copolymerized burnt pin hole 10 and are arranged with slit 11 in Raman
6 rear of optical filter is copolymerized burnt pin hole 10 for stopping under confocal mode for projecting beam splitting chip 12 after passing through laser
Spurious signal improves the three-dimensional space resolution ratio that sample is imaged, and sample signal intensity is promoted under non-confocal mode, beam splitting chip 12 is used
After making a part of Raman signal penetrate beam splitting chip 12, is read after the first narrow band filter 13 by the first detector 14, make to remain
After remaining Raman signal is reflected by beam splitting chip 12, read after the second narrow band filter 15 by the second detector 16.Beam splitting chip 12
For 10:90 beam splitting chip, after 90% Raman signal penetrates beam splitting chip 12, by the first detector 14 after the first narrow band filter 13
It reads, after 10% Raman signal is reflected by beam splitting chip 12, is read after the second narrow band filter 15 by the second detector 16.
The optional working range of beam splitting chip 12 is 400-700nm, and the first narrow band filter 13 selects 600/14nm, i.e., middle cardiac wave
Optical transmittance is greater than 93% within the scope of long 600nm, 593-607nm, to read the C-D Raman scattering within the scope of 588-608nm
Signal and the region Raman background signal；Second narrow band filter 15 selects 591/6 nm, i.e. central wavelength 591nm, 588-
Optical transmittance is greater than 93% within the scope of 595nm, to read the Raman background signal within the scope of 586- 596nm.
The Raman scattering Quick Acquisition of fixed wave length and the application of imaging device, comprising the following steps: in experimentation, swash
It shines and is issued through laser 1, filter out miscellaneous line by laser narrow-band optical filter 2, then after 3 adjusting strength of laser power attenuator,
Incident angle after being optimized laser by reflection microscope group is projected to Raman optical filter 6；Laser through Raman optical filter 6 reflection after by
Object lens 7 focus on the sample on glass slide 8；Auto-translating platform 9 can automatically move sample position according to software set, to reach
To scanning imagery purpose；Raman is returned to after being collected by the Raman scattering signal and Rayleigh scattering signal of sample scattering by object lens 7 to filter
Mating plate 6, is blocked in this Rayleigh scattering signal and Raman scattering signal penetrates Raman optical filter 6；Raman scattering signal using
It is copolymerized burnt pin hole 10 and slit 11 projects on beam splitting chip 12, in this 90% Raman signal by beam splitting chip again through the first narrowband
Optical filter 13, transmission range in C-D Raman wavelength region, after read by the first detector 14,10% Raman signal will be excessive
Beam piece is again through the second narrow band filter 15, Raman background signal area of the transmission range before C-D wavelength region, and C-D is drawn
Graceful wavelength region transmitance be 0, after read by the second detector 16；First detector 14 reads C-D Raman signal and Raman
Background signal, i.e. C curve in Fig. 3, the only Raman background signal that the second detector 16 is read, i.e. B curve in Fig. 3；Finally by
Algorithm deducts the background signal that the first detector 14 is read, to obtain pure C-D Raman signal.
Embodiment 2
Absorb the single cell Raman spectrum acquisition of the Escherichia coli of deuterium:
E.coli (deposit number ATCC25922, be purchased from ATCC) monoclonal is picked from the plate, 5ml LB is inoculated into
In fluid nutrient medium, it is placed in constant incubator (37 DEG C, 150rpm) and is incubated overnight.The bacterium that will be incubated overnight with 1:1000 ratio
It is transferred to the D that 5ml contains the various concentration that concentration range is 0% to 50%2In the LB liquid medium of O, constant incubator
4h is incubated in (37 DEG C, 150rpm).1ml bacterium solution is respectively taken, 5000rpm centrifugation 2min removes supernatant, 1ml sterile water, liquid relief is added
It robs piping and druming 3~5 times, 5000rpm centrifugation 2min removes supernatant, repeats above-mentioned add water wash step 1 time.Finally plus 1ml is sterile
Water, liquid-transfering gun piping and druming, which mixes bacterium solution, can carry out point sample detection.It when point sample, takes on 1 μ l sample to calcirm-fluoride glass slide, room temperature is certainly
So air-dry.
It excellent glass slide will be put takes under the burnt microscopic Raman system of copolymerization (Lab RAM HR, Horiba) and detected,
Found under 100 times of object lens E.coli it is unicellular and focus it is clear, carry out single cell Raman spectrum acquisition.Test condition is
532nm laser (power 100mW), grating: 300gr/mm, acquisition time: 20s.Obtain unicellular Raman light as shown in Figure 2
Spectrum.
Embodiment 3
Using the Raman scattering Quick Acquisition and imaging device of the fixed wave length of the utility model, laser 1 selects power
Adjustable 532nm laser, laser narrow-band optical filter 2 select 532/2nm, i.e. transmitance is greater than 90% at 532nm, transmittance curve
Full width at half maximum 2nm, to filter out the miscellaneous line in laser.
Beam splitting chip 12 is 10:90 beam splitting chip, after 90% Raman signal penetrates beam splitting chip 12, after the first narrow band filter 13
It is read by the first detector 14, after 10% Raman signal is reflected by beam splitting chip 12, by second after the second narrow band filter 15
Detector 16 is read.
12 working range of beam splitting chip is 400-1000nm, and the first narrow band filter 13 selects 600/14nm, i.e. central wavelength
Optical transmittance is greater than 93% within the scope of 600nm, 593-607nm, to read the letter of the C-D Raman scattering within the scope of 588-608nm
Number and the region Raman background signal；Second narrow band filter 15 selects 591/6 nm, i.e. central wavelength 591nm, 588-595nm
Optical transmittance is greater than 93% in range, to read the Raman background signal within the scope of 586- 596nm.
It is placed on calcirm-fluoride glass slide by the Escherichia coli ATCC25922 of deuterium-labeled (see embodiment 2), with the system acquisition
Its Raman spectrum.Raman scattering signal projects on beam splitting chip 12, in this 90% Raman signal that beam splitting chip is narrow through first again
Band optical filter 13, transmission range collect the Raman scattering signal of C curve ranges in Fig. 3 in C-D Raman wavelength region.10%
Raman signal by beam splitting chip again through the second narrow band filter 15, collect the Raman scattering signal of B curve ranges in figure.
To obtain the target Raman signal that target interval removes background Raman signal, carry out as follows:
1. correcting the reading of two groups (optical filter+detector) with standard sources；
2. measuring the Raman signal without deuterium-labeled Escherichia coli, the function calculated between two detector readings is closed
System: y=ax+b, wherein y is the reading of detector 16, and x is the reading of detector 15.By enough measured values, find quasi-
Close out a for most approaching true value, b parameter.
3. measuring the Raman signal of the Escherichia coli containing deuterium, the reading of detector 16 is y ', and the reading of detector 15 is x ', then is wanted
The echo signal value for the removal background asked are as follows: y=y '-(ax '+b)
It can understand the above description of the embodiments is intended to facilitate those skilled in the art and use practical
It is novel.Person skilled in the art obviously easily can make various modifications to these embodiments, and illustrating herein
General Principle be applied in other embodiments without having to go through creative labor.Therefore, the utility model is not limited to above-mentioned
Embodiment, those skilled in the art's announcement according to the present utility model, do not depart from improvement that the utility model scope is made and
Modification should be all within the protection scope of the utility model.
Claims (15)
1. the Raman scattering Quick Acquisition and imaging device of a kind of fixed wave length characterized by comprising
Raman scattering signal filter element: for making the Raman scattering signal of sample generation under laser action by wavelength filter device
Or the filtering of equipment selectivity,
Raman scattering signal acquisition unit: it for receiving the Raman scattering signal for passing through Raman scattering signal filter element, uses
Photon reading device or equipment are as detector.
2. the Raman scattering Quick Acquisition and imaging device of a kind of fixed wave length according to claim 1, which is characterized in that
Wavelength filter device or equipment used in Raman scattering signal filter element are one or more optical filters.
3. the Raman scattering Quick Acquisition and imaging device of a kind of fixed wave length according to claim 1, which is characterized in that
The photon reading device of Raman scattering signal acquisition unit or the detector of equipment are one or more photon counters or photoelectricity
Multiplier tube.
4. the Raman scattering Quick Acquisition and imaging device of a kind of fixed wave length according to claim 1, which is characterized in that
The Raman scattering signal filter element includes beam splitting chip (12), the first narrow band filter (13), the second narrow band filter (15),
After the beam splitting chip (12) is used to that a part of Raman signal to be made to penetrate beam splitting chip (12), through the first narrow band filter (13) mistake
Filter；After remaining Raman signal is reflected by beam splitting chip (12), filtered after the second narrow band filter (15).
5. the Raman scattering Quick Acquisition and imaging device of a kind of fixed wave length according to claim 4, which is characterized in that
The Raman scattering signal filtered after the first narrow band filter (13) is read by the first detector (14), through the second narrow band filter
(15) Raman scattering signal filtered afterwards is read by the second detector (16).
6. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 4 or 5 exist
In the beam splitting chip (12) is a:b beam splitting chip, and a and b are respectively the light after light is reflected transmitted through beam splitting chip and by beam splitting chip
By force, the ratio of a:b is greater than 0, less than 1.
7. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 4 or 5 exist
In beam splitting chip (12) working range at least covers any 100cm longer than laser (1) excitation wavelength-1The company of wave number
Continuous wave-length coverage section.
8. the Raman scattering Quick Acquisition and imaging device of a kind of fixed wave length according to claim 7, which is characterized in that
Beam splitting chip (12) working range is at least covered than laser (1) excitation wavelength red shift 2000cm-1To 2300cm-1Wave number model
The wavelength enclosed.
9. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 4 or 5 exist
In first narrow band filter (13) is selected in laser (1) excitation wavelength red shift direction 2040cm-1To 2300cm-1Wave number
Optical transmittance in range is greater than 93% optical filter.
10. the Raman scattering Quick Acquisition and imaging device of a kind of fixed wave length according to claim 4 or 5, feature
It is, second narrow band filter (15) is selected in laser (1) excitation wavelength red shift direction 1800cm-1-2040cm-1Or
2260cm-1To 2700cm-1The 30cm of middle arbitrary continuation in wave-number range-1Or 30cm-1Optics in above wave-number range penetrates
Rate is greater than 93% optical filter.
11. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 1 exist
In further including laser emission element, the laser emission element includes laser (1), laser narrow-band optical filter (2), described to swash
Light device (1) is used to filter out the miscellaneous line in laser for generating laser, the laser narrow-band optical filter (2).
12. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 11 exist
In, the laser emission element further includes laser power attenuator (3), for carrying out different degrees of decaying to laser intensity,
To adjust laser intensity.
13. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 10 exist
In the laser (1) is 488nm laser, 514nm laser, 532nm laser, 633nm laser or 785nm laser
One of device.
14. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 1 exist
In further including Raman optical filter (6) and object lens (7), the Raman optical filter (6) will be for that will reflect the laser that microscope group launches
It is focused on sample after reflection by object lens (7), the object lens (7) are used to collect the Raman scattering signal of sample scattering and Rayleigh dissipates
Signal is penetrated, and Raman scattering signal and Rayleigh scattering signal is made to return to Raman optical filter (6)；
The Rayleigh scattering signal that the Raman optical filter (6) is also used to that sample is stopped to scatter, through the Raman scattering of sample scattering
Signal.
15. the Raman scattering Quick Acquisition and imaging device, feature of a kind of fixed wave length according to claim 14 exist
In, the Raman scattering signal acquisition unit includes being copolymerized burnt pin hole (10), slit (11),
The burnt pin hole (10) of the copolymerization and slit (11) are arranged at Raman optical filter (6) rear, for projecting after passing through laser
To on beam splitting chip (12),
The burnt pin hole (10) of the copolymerization under confocal mode for stopping spurious signal, and promotion sample signal is strong under non-confocal mode
Degree.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821011459.1U CN208736797U (en) | 2018-06-28 | 2018-06-28 | The Raman scattering Quick Acquisition and imaging device of fixed wave length |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821011459.1U CN208736797U (en) | 2018-06-28 | 2018-06-28 | The Raman scattering Quick Acquisition and imaging device of fixed wave length |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN208736797U true CN208736797U (en) | 2019-04-12 |
Family
ID=66026273
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201821011459.1U Active CN208736797U (en) | 2018-06-28 | 2018-06-28 | The Raman scattering Quick Acquisition and imaging device of fixed wave length |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN208736797U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108627495A (en) * | 2018-06-28 | 2018-10-09 | 上海氘峰医疗器械有限公司 | The Raman scattering Quick Acquisition and imaging device of fixed wave length |
| CN114113213A (en) * | 2020-08-28 | 2022-03-01 | 西门子股份公司 | Measuring device for determining the gross or net heating value of a hydrocarbon-containing fuel gas |
-
2018
- 2018-06-28 CN CN201821011459.1U patent/CN208736797U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108627495A (en) * | 2018-06-28 | 2018-10-09 | 上海氘峰医疗器械有限公司 | The Raman scattering Quick Acquisition and imaging device of fixed wave length |
| CN114113213A (en) * | 2020-08-28 | 2022-03-01 | 西门子股份公司 | Measuring device for determining the gross or net heating value of a hydrocarbon-containing fuel gas |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108627495A (en) | The Raman scattering Quick Acquisition and imaging device of fixed wave length | |
| Bellon et al. | Feasibility and performances of a new, multiplexed, fast and low-cost fiber-optic NIR spectrometer for the on-line measurement of sugar in fruits | |
| US7154599B2 (en) | Spectrometer incorporating signal matched filtering | |
| US4560881A (en) | Method and apparatus for enhanced detection of electromagnetic signals | |
| US10267734B2 (en) | Wavelength dispersive microscope spectrofluorometer for characterizing multiple particles simultaneously | |
| CA2937696C (en) | Method of spectroscopic analysis of a diamond and apparatus thereof | |
| JP2019513236A (en) | Cell sorting using a high-throughput fluorescence flow cytometer | |
| CN101118208A (en) | Optical system for a particle analyzer and particle analyzer using same | |
| CN1898549B (en) | Precise flow-oriented multi-angle remission sensor | |
| CA2065668A1 (en) | Spectroscopically correlated light scanning microscopy | |
| WO1998022802A1 (en) | Method and device for combined absorption and reflectance spectroscopy | |
| US7999936B1 (en) | Combined transmittance and angle selective scattering measurement of fluid suspended particles for simultaneous determination of refractive index, extinction coefficient, particle size and particle density | |
| US3598994A (en) | Method and apparatus for sensing fluorescent substances | |
| CN101995387A (en) | Multi-functional ultraviolet-visible spectrometer | |
| CN208736797U (en) | The Raman scattering Quick Acquisition and imaging device of fixed wave length | |
| CN107478550A (en) | The triple channel emerging system of real-time detection of particles size and property | |
| JPH10253624A (en) | Particle measuring device | |
| CN206557053U (en) | A kind of oil quality detection means | |
| CN100526883C (en) | Reflection photometer for gold standard immunoassay strips | |
| CN103575712A (en) | Particle fluorescence detection wavelength instant configuration beam splitting system | |
| MXPA98000206A (en) | Identification of recirculable carpet materials using an infrared man spectrometer | |
| Desiderio et al. | Microstructure profiles of laser-induced chlorophyll fluorescence spectra: evaluation of backscatter and forward-scatter fiber-optic sensors | |
| CN212844874U (en) | Liquid measurement system based on optical cavity enhancement | |
| CN209961688U (en) | Rapid detection instrument for pathogenic microorganism drug sensitivity | |
| CN109100021A (en) | A kind of time-resolved spectrum and lifetime measurement module and device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210628 Address after: 201802 room 108, 1st floor, building 18, No. 1188, Huyi highway, Jiading District, Shanghai Patentee after: Shanghai deuterium peak Medical Technology Co.,Ltd. Address before: 201802 J1953 1, 12 floor, B 1, 399 Ren Ren Road, Jiading District, Shanghai Patentee before: SHANGHAI D-BAND MEDICAL INSTRUMENT Co.,Ltd. |